IL112249A - Pharmaceutical compositions containing di and tricyclic pyrimidine derivatives for inhibiting tyrosine kinases of the epidermal growth factor receptor family and some new such compounds - Google Patents

Abstract

A pharmaceutical composition adapted for administration as an inhibitor of the epidermal growth factor receptor family of tyrosine kinases, comprising a therapeutically effective amount of a compound of the formula 2933 י' בכסלו התשס" ב - November 25, 2001 or a pharmaceutical salt or hydrate thereof, in admixture with a pharmaceutically acceptable excipient, diluent or carrier: X= O, S or NH; n = 0, 1, 2; R1 = H or alkyl (1-4 carbon atoms); if n = 2, R1 can be independently H or alkyl (1-4 carbon atoms) on either linking carbon atom; R2 is alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms), halo (fluoro, chloro, bromo, iodo), perfluoroalkyl (1-4 carbon atoms), hydroxy, acyloxy of the formula -O-C(O)R where R is alkyl of 1-4 carbon atoms or cycloalkyl of 3-8 carbon atoms, amino, mono or dialkylamino (1-4 carbon atoms in each alkyl group), mono or dicycloalkylamino (3-8 carbon atoms in each ring), hydroxymethyl, acyl of the formula -C(O)R, cyano, alkylthio (1-4 carbon atoms), alkylsulfinyl (1-4 carbon atoms), alkylsulfonyl (1-4 carbon atoms), cycloalkylthio (3-8 carbon atoms), cycloalkylsulfinyl (3-8 carbon atoms), cycloalkylsulfonyl, (3-8 carbon atoms), sulfamoyl, mono or dialkylsulfamoyl (1-4 carbon atoms in each alkyl group), mono or dicycloalkylsulfamoyl (3-8 carbon atoms in each ring), mercapto, carboxy, carbamoyl (-C(O)-NH2), mono or dialkylcarbamoyl (1-4 carbon atoms in each alkyl group), mono or 2934 י' בכסלו התשס" ב - November 25, 2001 dicycloalkylcarbamoyl (3-8 carbon atoms in each ring), alkoxycarbonyl (1-4 carbon atoms in the alkyl moiety), cycloalkoxycarbonyl (3-8 carbon atoms in the ring), alkenyl (2-4 carbon atoms), cycloalkenyl (4-8 carbon atoms), alkynyl (2-4 carbon atoms), or two R2 taken together on contiguous carbon atoms can form a carbocyclic ring of 5-7 members; m=0-3, wherein Ar is phenyl, thienyl, furanyl, pyrrolyl, pyrimidyl, imidazolyl, pyrazinyl, oxazolyl, thiazolyl, naphthyl, benzothienyl, benzofuranyl, indolyl, quinolinyl, isoquinolinyl and quinazolinyl; at least one of A-E cannot be carbon, unless R3 and R4 taken together form an aromatic or heteroaromatic ring, as defined below; any one of the A-E can be N with proviso that if R3 and R4 are either a lone pair on N or H, then n=0 or 1; both B and E can be N, with the following provisos: the other two atoms of B-C in that ring are C, R3 is not H, and n is 0 or 1; any two contiguous positions in A-E can be a single heteroatom, N, O or S, wherein one of the two remaining named positions of A-E must be C(H), and the other can be C(H) or N, resulting in a 5 membered fused ring; R3 and R4 are independently H, alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms, hydroxy, acyloxy (1-4 carbon atoms), amino, mono or dialkylamino (1-4 carbon atoms), mono or dicycloalkylamino (3-8 carbon atoms), carbonato (-OC(O)OR), where R is alkyl (1-4 carbon atoms) or cycloalkyl (3-8 carbon atoms), mono or dialkyl substituted ureido, thioureido, or N- or O-linked urethano (1-4 carbon atoms in each alkyl group), alkylthio (1-4 carbon atoms) or cycloalkylthio (3-8 carbon atoms), mercapto, alkenyl (2-4 carbon atoms), hydrazino, N(-lower alkylhydrazino (1-4 carbon atoms), acylamino (1-4 carbon atoms), hydroxylamino, O-alkylhydroxylamino (1-4 carbon atoms), or taken together can be methylene-, ethylene -or propylenedioxy, or taken together form a fused pyrrolidine, tetrahydrofuran, piperidine, piperazine, morpholine or thiomorpholine ring, or taken together form a further aromatic ring selected from the group consisting of phenyl, furan, thiophene, pyrazole, isoxazole, isothiazole, pyrrole, oxazole, imidazole and thiazole, optionally substituted with 1-3 of the R2 substituents; if B and D are C, A and E can be N, provided that at least one of R3 and R4 is lower alkoxy, or the two taken together form an aromatic ring sele

Description

112249/3 PHARMACEUTICAL COMPOSITIONS CONTAINING DI AND TRICYCLIC PYRIMIDINE DERIVATIVES FOR INHIBITING TYROSINE KINASES OF THE EPIDERMAL GROWTH FACTOR RECEPTOR FAMILY AND SOME NEW SUCH COMPOUNDS WARNER-LAMBERT COMPANY C: 31317 112249/3 -1 - PHARMACEUTICAL COMPOSITIONS CONTAINING DI AND TRICYCLIC PYRIMIDINE DERIVATIVES FOR INHIBITING TYROSINE KINASES OF THE EPIDERMAL GROWTH FACTOR RECEPTOR FAMILY AND SOME NEW SUCH COMPOUNDS Technical Field The present invention relates to di and tricyclic heteroaromatic compounds which inhibit the epidermal growth factor receptor and related receptors and, in particular, their tyrosine kinase enzymic activity.

Background Art Cancer is generally a disease of the intracellular signalling system, or signal transduction mechanism. Cells receive instructions from many extracellular sources, instructing them to either proliferate or not to proliferate. The purpose of the signal transduction system is to receive these and other signals at the cell surface, get them into the cell, and then pass the signals on to the nucleus, the cytoskeleton, and transport and protein synthesis machinery. The most common cause of cancer is a series of defects, either in these proteins, when they WLC 0131 PUS -2- are mutated, or in the regulation of the quantity of the protein in the cell such that it is over or under produced. Most often, there are key lesions in the cell which lead to a constitutive state whereby the cell nucleus receives a signal to proliferate, when this signal is not actually present. This can occur through a variety of mechanisms. Sometimes the cell may start to produce an authentic growth factor for its own receptors when it should not, the so-called autocrine loop mechanism. Mutations to the cell surface receptors, which usually signal into the cell by means of tyrosine kinases, can lead to activation of the kinase in the. absence of ligand, and passing of a signal which is riot really there. Alternatively, many surface kinases can be overexpressed on the cell surface leading to an inappropriately strong response to a weak signal. There are many levels inside the cell at which mutation or overexpression can lead to the same spurious signal arising in the cell, and there are many other kinds of signalling defect involved in cancer. This invention touches upon cancers which are driven by the three mechanisms just described, and which involve cell surface receptors of the epidermal growth factor receptor tyrosine kinase family (EGFR) . This family consists of the EGF receptor (also known as Erb-Bl) , the Erb-B2 receptor, and its constituitively active oncoprotein mutant Neu, the Erb-B3 receptor and the Erb-B4 receptor.

Additionally, other biological processes driven through members of the EGF family of receptors can also be treated by compounds of the invention described below.

The EGFR has as its two most important ligands Epidermal Growth Factor (EGF) and Transforming Growth Factor alpha (TGFalpha) . The receptors appear to have only minor functions in adult humans, but are apparently implicated in the disease process of a large portion of all cancers, especially colon and breast cancer. The closely related Erb-B2, Erb-B3 and Erb-B4 receptors have a family of Heregulins as their major ligands, and receptor overexpression and mutation have been unequivocally demonstrated as the major risk factor in poor prognosis breast cancer. Additionally, it has been demonstrated that all four of the members of this family of receptors can form heterodimeric signalling complexes with other members of the family, and that this can lead to synergistic transforming capacity if more than one member of the family is overexpressed in a malignancy. Overexpression of more than one family member has been shown to be relatively common in human malignancies.

The proliferative skin disease psoriasis has no good cure at present. It is often treated by anticancer agents such as methotrexate, which have very serious side effects, and which are not very effective at the toxicity-limited doses which have to be used. It is believed that TGFalpha is the major growth factor overproduced in psoriasis, since 50% of transgenic mice which overexpress TGF alpha develop psoriasis. This suggests that a good inhibitor of EGFR signalling could be used as an antipsoriatic agent, preferably, but not necessarily, by topical dosing.

WLC 0131 PUS -4- EGF is a potent mitogen for renal tubule cells. Fourfold increases in both EGF urinary secretion and EGF mRNA have been noted in mice with early stage streptozoicin- induced diabetes. In addition increased expression of the EGFR has been noted in patients with proliferative glomerulonephritis (Roychaudhury et al. Pathology 1993, 25, 327) . The compounds of the current invention should be useful in treating both proliferative glomerulonephritis and diabetes-induced renal disease.

Chronic pancreatitis in patients has been reported to correlate with large increases in expression for both EGFR and TGF alpha. (Korc et al . Gut 1994, 35, 1468) . In patients showing a more severe form of the disease, typified by an enlargement of the head of the pancreas, there was also shown to be overexpression of the erb-B2 receptor (Friess et al. Ann. Surg. 1994, 220, 183). The compounds of the current invention should prove useful in the treatment of pancreatitis.

In the processes of blastocyte maturation, blastocyte implantation into the uterine endometrium, and other periimplantation events, uterine tissues produce EGF and TGF alpha (Taga Nippon Sanka Fujinka Gakkai Zasshi 1992, 44, 939), have elevated levels of EGFR (Brown et al. Endocrinology, 1989, 124, 2882), and may well be induced to produce heparin-binding EGF by the proximity of the developing, but not arrested, blastocyte (Das et al . Development 1994, 120, 1071) . In turn the blastocyte has quite a high level of TGF alpha and EGFR expression (Adamson Mol . Reprod. Dev. - - 1990, 27, 16). Surgical removal of the submandibular glands, the major site of EGF secretion in the body, and treatment with anti-EGFR monoclonal antibodies both greatly reduce fertility in mice (Tsutsumi et al. J. Endocrinology 1993, 138, 437), by reducing successful blastocyte implantation. Therefore, compounds of the current invention should prove to have useful contraceptive properties.

PCT published patent application Nos. WO92/07844 published May 14, 1992 and WO92/14716 published September 3, 1992 describe 2.4-diaminoquinazoline as potentiators of chemotherapeutic agents in the treatment of cancer.

PCT published patent application No. WO92/20642 published November 26, 1992 discloses bismono- and bicyclic aryl and heteroaryl compounds which inhibit EGF and/or PDGF receptor tyrosine kinase.

It is an object of the present invention to mhibit the mitogenic effects of epidermal growth factor utilizing an effective amount of di or tricyclic pyrimidine derivatives, in particular fused heterocyclic pyrimidine derivatives.

It is another object of the present invention to provide di and tricyclic pyrimidine derivatives, in particular fused heterocyclic pyrimidine derivatives, as inhibitors of the EGF, Erb-B2 and Erb-B4 receptor tyrosine kinases.

It is yet another object of the present invention to provide di and tricyclic pyrimidine derivatives, 1 12249/4 -6-in particular fused heterocyclic pyrimidine derivatives, that are useful at low dosages as inhibitors of EGF-induced mitogenesis . This therefore leads to a further object of compounds having extremely low cytotoxicity.

It is a further object of the present invention to provide bicyclic pyrimidine derivatives, in particular fused heterocyclic pyrimidine derivatives, that are useful in suppressing tumors, especially breast cancers, where mitogenesis is ..heavily driven by EGFR family members.

It is another object of the present invention to provide bicyclic pyrimidine derivatives, in particular fused heterocyclic pyrimidine derivatives, that have utility as chronic therapy as inhibitors of EGF-induced responses.

It is another object of the current invention to provide bicyclic pyrimidine derivatives, in particular fused heterocyclic pyrimidine derivatives, that have utility as therapeutic agents against proliferative overgrowth diseases, including but not limited to, synovial pannus invasion in arthritis, vascular restenosis, psoriasis and angiogenesis . The compounds disclosed herein also are useful to treat pancreatitis and kidney disease and as contraceptive agents.

NOTICE UNDER REGISTRAR'S CIRCULAR NO. (P)23 Portions of the present specification that fall outside the scope of the claims do not relate directly to the present invention. This Notice is not meant to disclaim any legitimate rights to which the Patentee is legally entitled, especially any rights in accordance with Section 49 of the Law.

Summary of the Invention The present invention provides 112249/4 -7- a pharmaceutical composition adapted for administration as an inhibitor of the epidermal growth factor receptor family of .tyrosine kinases, comprising a therapeutically effective amount of a compound of Formula I or a pharmaceutical salt or hydrate thereof, in admixture with a pharmaceutically acceptable excipient, diluent or carrier: Formula. I where : 112249/4 -8- X = o, s or NH; n = 0, 1, 2; R1 = H or alkyl (1-4 carbon atoms);- if n = 2, R1 can be independently H or alkyl {1-4 carbon atoms) on either linking carbon atom; R2 is alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms), halo (fluoro, chloro, bromo, iodo) , perfluoroalkyl (1-4 carbon atoms), hydroxy, acyloxy of the formula- -O-C(0)R where R is alkyl of 1-4 carbon atoms or cycloalkyl of 3-8 carbon atoms, amino, mono or dialkylamino (1-4 carbon atoms in each alkyl group) , mono or dicycloalkylamino (3-8 carbon atoms in each ring), hydroxymethyl , acyl of the formula -C(0)R, cyano, alkylthio (1-4 carbon atoms), alkylsulfinyl (1-4 carbon atoms), alkylsulfonyl (1-4 carbon atoms), cycloalkylthio (3-8 carbon atoms), cycloalkylsulfiiiyl (3-8 carbon atoms) , cycloalkylsulfonyl (3-8 carbon atoms), sulfamoyl , mono or dialkylsulfamoyl (1-4 carbon atoms in each alkyl group) , mono or dicycloalkylsulfamoyl (3-8 carbon atoms in each ring), mercapto, carboxy, carbamoyl (-C(0)-NH2), mono or dialkylcarbamoyl (1-4 carbon atoms in each alkyl group) , mono or dicycloalkyl- carbamoyl (3-8 carbon atoms in each ring), alkoxycarbonyl (1-4 carbon atoms in the alkyl moiety), cycloalkoxy- carbonyl (3-8 carbon atoms in the ring), al'kenyl (2-4 carbon atoms), cycloalkenyl (4-8 carbon atoms), alkynyl (2-4 carbon atoms), or two R2 taken together on contiguous carbon atoms can form a carbocyclic ring of 5-7 members; : 1 -9- m = 0-3, wherein Ar is phenyl, tlueoyl, ihranyi, pyrrolyi, pyrimidyl, imidazole ' pyrazmyl, oxazoly], ftiazdyi, nap thy], beiizotliieny], benzo&ranyl, mdolyl, qumolinyl, isoqmnolmyl and qmnazoiiny]; at least one of A-E cannot be carbon, unless R3 and R4 taken together form an aromatic or heteroaro- matic ring, as defined below; any one of the A-E can be N with proviso that if R3 and R4 are either a lone pair on N or H, then n=0 or 1 ; both B and E can be N, with the following provisos: the other two atoms of A-E in that ring are C, R4 is not- H, and n is 0 or 1; both A and D can be N, with the following provisos: the other two atoms of B-C in that ring are C, R3 is not H, and n is 0 or 1; any two contiguous positions in A-E can be a single heteroatom, N, O or S, wherein one of the two remaining named positions of A-E must be C(H), and the other can be C(H) or N, resulting in a 5 membered fused ring ; R3 and R4 are independently H, alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy (1-4 carbon atoms) , cycloalkoxy (3-8 carbon atoms) , hydroxy, acyloxy (1-4 carbon atoms) , amino, mono or dialkylamino (1-4 carbon atoms) , mono or dicycloalkylamino (3-8 carbon atoms), carbonato (-OC(O)OR), where R is alkyl (1-4 carbon atoms) or cycloalkyl (3-8 carbon atoms), mono or dialkyl substituted ureido, thioureido, or N- or O-linked urethano (1-4 carbon atoms in each alkyl group), alkylthio (1-4 carbon atoms) or cycloalkylthio (3-8 carbon atoms), mercapto, alkenyl (2-4 carbon atoms), hydrazino, N' -lower alkylhydrazino (1-4 carbon atoms) , acylamino (1-4 carbon atoms) , hydroxy1amino, .0-alkylhydroxylamino (1-4 carbon atoms) , or taken together can be methylene-, ethylene- or propylenedioxy, or taken together form a fused pyrrolidine, tetrahydrofuran, . piperidine, piperazine, morpholine or thiomorpholine ring, or taken together form a further aromatic ring selected from the group consisting of phenyl, furan, thiophene, pyrazole, isoxazole, isothiazole, pyrrole, oxazole, imidazole and thiazole, optionally substituted with 1-3 Of the R2 substituents; if B and D are C, A and E can be N, provided that at least one of R3 and R4 is lower alkoxy, or the two taken together form an aromatic ring selected from the group consisting of phenyl, furan, thiophene, pyrrole,, oxazole, imidazole and thiazole, optionally substituted with 1-3 of the R2 subs ituents; and if one or two of A through E are N, then if R3 or R4 is on a neighboring C atom to one of the N atoms, the named substituent cannot be either OH or SH; R5 and R6 are H when A and/or E are a carbon atom; if any of the substitutents R R2, R3 or R4 contain chiral centers, or in the case wherein ^creates chiral centers on the linking atoms, then all stereoisomers thereof both separately and as racemic and/or diastereoisomeric mixtures are included.

This invention further provides a compound of Formula I; Formula I a pharmaceutical salt or hydrate ■ thereofr where : X = 0, S or NH "' n = 0, 1, 2; Rl = H or alkyl (1-4 carbon atoms); if n = 2, can be independently H or alkyl (1-4 carbon atoms) either linking carbon atom; -10a- R? is alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms), halo (fluoro, chloro, bromo, iodo), perfluoroalkyl (1-4 carbon atoms), hydroxy, acyloxy of the formula; -0-C(0)R where B is alkyl of 1-carbon atoms or cycloalkyl of 3-8 carbon atoms, amino, mono or dialkylaraino (1-4 carbon atoms in each alkyl group), mono or dicycloalkylamino (3-8 carbon atoms in each ring), hydroxymethyl , acyl of the formula -C(0)R, cyano, alkylthio (1-4 carbon atoms), alkylsulfinyl (1-4 carbon atoms), alkylsulfonyl (1-4 carbon atoms), cycloalkylthio (3-8 carbon atoms), cyclodkylsulfiayl (3-8 carbon atoms), cycloalkylsulfonyl (3-8 carbon atoms) , sulfamoyl , mono or dialkylsulfamoyl (1-4 carbon atoins in each alkyl group), mono or dicycloalkylsulfamoyl (3-8 carbon atoms in each ring), snercanto carboxy, carbamoyl (-C(0)-NH2), mono or diaIkylcarbamoyl (1-4 carbon atoms in each alkyl group), mono or dicycloalkyl-carbamoyl (3-8 carbon atoms in each ring), alkox ca bon l (1-4 carbon atoms in the alkyl moiety), cycloaikoxy-carbonyl (3-8 carbon atoms in the ring) , ' alkenyl (2-4 carbon atoms), cycloalkenyl (4-8 carbon atoms), alkynyl (2-4 carbon atoms), or two R2 taken together on contiguous carbon atoms can form a carbocyclic ring of 5-7 members; and m - 0-3, wherein Ar is phenyl, thienyl, furanyl, pyrrolyl, pyrimidyl, imidazolyl, pyrazinyl, oxazolyl , thiazolyl, naphthyl, benzothienyl , benzo-furanyl , indolyl, quinolinyl, isoquinolinyl and quina-zolinyl ,- at least one of A-E cannot be carbon, unless R3 and R4 taken together form an aromatic or heteroaro-matic ring, as defined below ■ any one of the A-E can be N with proviso that if R3 and R"* are either a lone pair on N or H, then n=0 or 1; -lob- both B and E can be N, with the following PrOV41SCS: the other two atoms of A-E in that ring are C, R4 is not H, and n is 0 or i; both A and D can be N, with the following Pr0VisO :: the other tw° atoms of B-C in that ring are C, R3 is not H, and n is 0 or i; R3 and R4 are independently H, alkyl (1-4 carbon atoms), cycioalkyl (3-3 carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms), hydroxy, acyloxy (1-4 carbon atoms), amino, mono or dialkyla ino (1-4 carbon atoms) , mono or dicycloalkylamino (3-8 carbon atoms) , carbonato (-OC(O)OR), where R is alkyl (1-4 carbon atoms) or cycioalkyl (3-8 carbon atoms), mono or dialkyl substituted ureido, thioureido, or N- or O-linked urethano (1-4 carbon atoms in each alkyl group), alkylthio (1-4 carbon atoms) or cycloalkylthio (3-8 carbon atoms), mercapto, alkenyl (2-4 carbon atoms), hydrazine N' - alkylhydrazirio '{1-4 carbon atoms) , acylamino (1-4 carbon atoms) , hydroxyiamino, O-alkylhydroxylamino (1-4 carbon atoms) , or taken together can be methylene-,, ethylene- or propyienedioxy , or taken together form a fused pyrrolidine, tetrahydrofuran, piperidine, piperazine, morpholine or thiomorpholine ring, or taken together form a further aromatic ring selected from the group consisting of phenyl, furan, thiophene, pyrazole, isoxa2ole, isothiazcie, pyrrole, oxazoj.e, imidazole and thiazole, optionally substituted with 1-3 of the R2 substituents ; if B and D are C, A and E can be N, provided that at least one of R3 and R4 is lower alkoxy, or the two taken together form an aromatic ring selecced from the group consisting of phenyl, furan, thiophene, pyrrole, oxazole, imidazole and thiazole, optionally substituted with 1-3 of the R2 substituents; and -10c- if one or two of A through E are N, then if R3 or R1 is on a neighboring C atom to one cf the N atoms, the named substituent cannot be either OH or SB; and R5 and R6 are H when A and/or E are a carbon atom; if any of the SObst tutents R\ R2, R3 orR4 contain - chiral centers , or in the case wherein R creates chiral centers on the linking atoms, then all stereoisomers thereof both separately and as racemic and/or diastereoisojneric mixtures are included.

Brief Description Of The Drawings FIGURE 1 shows an effect of compounds of Examples 6 and 7 on EGF receptor auiophosphorylation in. A431 human, epidermoid carcinoma; -Π- FIGURE 2 shows an effect of compound of Example 8 on EGF receptor autophosphorylation in A431 human epidermoid carcinoma; FIGURE 3 shows a time course for the inhibition of EGF receptor autophosphorylation in A431 by compound of Example 27; FIGURE 4 shows an effect of compound of Example 27 on EGF receptor autophosphorylation in A431 cells; FIGURE 5 shows inhibition of EGF receptor autophosphorylation in A431 human epidermoid carcinoma by compound of Example 40; FIGURE 6 shows an effect of compound of Example 40 on growth factor-mediated tyrosine phosphorylation in Swiss 3T3; FIGURE 7 shows an effect of compound of Example 40 on growth factor dependent expression of c-jun mRNA in Swiss 3T3 mouse fibroblasts; FIGURE 8 shows an effect of compound of Example 40 on growth factor mediated expression of p39c jim; FIGURE 9 shows an effect of compound of Example 59 of EGF receptor autophosphorylation in A431 human epidermoid carcinoma; FIGURE 10 shows an effect of compound of Example 60 on EGF receptor autophosphorylation in A431 human epidermoid carcinoma; -12- FIGURE 11 shows an effect of compound of Example 61 on EGF receptor autophosphorylation in A431 human epidermoid carcinoma; FIGURE 12 shows an effect of compound of Example 70 on EGF receptor autophosphorylation in A431 human epidermoid carcinoma; FIGURE 13 is a chart showing an inhibition of EGF receptor tyrosine kinase by compound of Example 27; FIGURE 14 is a graph showing an effect of compound of Example 40 on growth factor-mediated mitogenesis in Swiss 3T3 murine fibroblasts; FIGURE 15 is a photograph of an IH 3T3 mouse fibroblast line, transfected with the human EGFR gene showing a normal flattened morphology; FIGURE 16 is a photograph of the same cell line treated with 100 ng mL of EGF showing a typical spindly transformed morphology; and FIGURE 17 is a photograph of the same cell line in the presence of both 100 ng/mL of EGF and 5 μπι of compound of Example 27 showing the morphology reverted from the transformed type back to the normal type.

Description of Preferred Embodiments 1. A preferred form of the invention has X = NH, n = 0 or 1, in which case R1 - H, the aromatic ring phenyl optionally substituted, B, D & E carbon, -13- with A nitrogen and R3 or R4 H, with the other one alkoxy or halogen. 2. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 or R4 H, with the other one amino. 3. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 or R4 H, with the other one mono or dialkylamino .

. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 or R4 H, with the other one hydrazine 5. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 or R4 H, with the other one alkyl. 6. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 and R4 alkoxy. 7. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the -14- aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 and R4 alkyl. 8. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen, and R3 or R4 amino, with the other one alkoxy. 9. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen, and R3 or R4 mono or dialkylamino, with the other one alkoxy. 10. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 mono or dialkylamino, with R4 hydroxy.

A suitable ring structure for groups 1-10 is: 11. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E -15- carbon, with A nitrogen, and R3 and R4 taken together are methyl enedioxy, ethylenedioxy, 2,3-fused piperazine, 2,3-fused morpholine or 2,3-fused thiomorpholine. Suitable ring structures are: 12. Another preferred form of the invention has X = H, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R4 alkoxy or halogen. 13. Another preferred form of the invention has X = ΝΉ, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R4 amino or acylamino. 14. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R4 mono or dialkylamino. 15. Another preferred form of the invention has X = NH, n = 0 or 1 , in which case R1 = H, the -16- aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R4 hydrazino. 16. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R4 alkyl.

A suitable ring structure for groups 12- 17. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 alkoxy or halogen. 18. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 amino. 19. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 mono or dialkylamino. -17- 20. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 hydrazine 21. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 alkyl.

A suitable ring structure for groups 17-21 is: 22. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or RA H, with the other one alkoxy. 23. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or R4 H, with the other one amino. 24. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the -18- aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or R4 H, with the other one mono or dialkylamino . 25. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or R4 H, with the other one hydrazine 26. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or R4 H, with the other one alkyl. 27. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 and R4 alkoxy. 28. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 and R4 alkyl. 29. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen, and R3 or R4 amino, with the other one alkoxy. 30. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the -19- aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen, and R3 or R4 mono or dialkylamino, with the other one alkoxy. 31. Another preferred form of the invention has X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R4 mono or dialkylamino, with R3 hydroxy.

A suitable ring structure for groups 22-31 is: 32. Another preferred form of the invention has X- NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen, and R3 and R4 taken together are methylenedioxy, ethylenedioxy, 2,3-fused piperazine, 2,3-fused morpholine or 2,3-fused tmomorpholine. 33. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 alkoxy. -20- 34. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 mono or dialkyla ino. 35. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 amino. 36. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 hydrazino .

A suitable ring structure for groups 33-36 is: 37. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbon, with A and E nitrogen and R3 and R4 alkoxy. 38. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbon, with A and E nitrogen and R3 and R4 mono or dialkylamino. - - 39. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbonf with A and E nitrogen and R3 or alkoxy, with the other mono or dialkylamino. 40. Another preferred form of the invention has X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbon, with A and E nitrogen and R3 and taken together are ethylenedioxy, 2,3-fused piperazine, 2,3-fused morpholine or 2,3-fused thiomorpholine .

A suitable ring structure for groups 37-40 is: 41. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a sulfur atom, with D & E carbon, or A & B are carbon with D and E taken together as a sulfur atom, with R"9 or R3 H, alkyl, alkoxy, amino, or mono or dialkylamino. 42. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the -22- aromatic ring phenyl optionally substituted/ and either A and B taken together are an oxygen atom, with D & E carbon, or A & B are carbon with D and E taken together as an oxygen atom, with R4 or R3 H, alkyl, alkoxy, amino, or mono or dialkylamino . 43. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a nitrogen atom, with D & E carbon, or A & B are carbon with D and E taken together as a nitrogen atom, with R4 or R3 H, alkyl, alkoxy, amino, or mono or dialkylamino . 44. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a sulfur atom with D carbon and E nitrogen, or D and E taken together are a sulfur atom, and A is nitrogen and B is carbon, with R3/4 H, alkyl, alkoxy, amino, or mono or dialkylamino. 45. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are an oxygen atom with D carbon and E nitrogen, or D and E taken together are an oxygen atom, and A is nitrogen and B is carbon, with R3 4 H, alkyl, alkoxy, amino, or mono or dialkylamino. -23- 46. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A and B taken together are a nitrogen atom, and D is carbon and E is nitrogen, with R3 6 H, or alkyl, and R4 H, alkyl, alkoxy, amino, or mono or dialkylamino . 47. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are an oxygen atom with D nitrogen and E carbon, or D and E taken together are an oxygen atom with B nitrogen and A carbon, with R3/6 H, alkyl, alkoxy, amino, or mono or dialkylamino. 48. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a sulfur atom with D nitrogen and E carbon, or D and E taken together are a sulfur atom with B nitrogen and A carbon, with R3 6 H, alkyl, alkoxy, amino, or mono or dialkylamino. 49. Another preferred form of the invention has X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a nitrogen atom with D nitrogen and E carbon, or D and E taken together are a nitrogen atom with B nitrogen and A carbon, with R3 6 H or alkyl if on nitrogen, or H, alkyl, alkoxy, amino, or mono or dialkylamino if on carbon.

Other suitable ring structures are: Where Z B nitrogen, oxygen or sulfur The compounds of the present invention are prepared according to a number of alternative reaction sequences .

Preparative Routes to Compounds of the Invention (The schemes are depicted graphically on pages 135-144) Scheme 1 - Route for Preferred Groups 1-5, R4 = H-See pages 12 and 13.

Displacement of the 2-chloro of 2, 6-dichloro-3-nitropyridine is carried out by cuprous cyanide in NMP. Displacement of the second chlorine of this nitrile by fluoride at this step can be advantageous. This is followed by a mild reduction of the nitro group, under conditions where the halogen is not hydrogenolysed. Hydrolysis of the nitrile followed by orthoformate cyclization, and Vilsmeier-type chlorination will give the dihalo-pyridopyrimidine . Displacement . of the more reactive 4-chlorine with an appropriate amine is followed by displacement of the 6-halogen with the appropriate nucleophile, ammonia, lower alkylamine, hydrazine, methoxide, to form the final products. (NMP is a solvent, N-methyl-2-pyrrolidone) . -25- Scheme 2 - Route to Preferred Groups 1-5, R3 = H-See pages 12 and 13 Displacement of chlorine f 2-chloro-3, 5-dinitropyridine is accomplished with CuCN -26- in NMP. Reduction of the nitro groups to amines is followed by hydrolysis of the nitrile to an amide. This is cyclized to the pyrimidone with orthoformate, which is converted to the chloride by P0C13 or possibly turned into the methylthio derivative by treatment with phosphorus pentasulfide followed by Mel and a mild base. Displacement with the appropriate amine gives the desired 7-amino compound. The amine functionality can be reductively alkylated or activated by diazotisation of the amino group under acidic or basic conditions, followed by a reduction to the hydrazide, or conversion into a lower alkyl ether, or to a halogen followed by a cuprate or Stille coupling by methods familiar to those skilled in the art. Alternatively, the amine can be reductively aminated, or acylated and reduced to form the alkylamino side chain.

Scheme 3 - Route to Preferred Groups 6 and 8-10 where R4 = RO-See pages 13 and 14.

The known metalation of 2, 6-difluoropyridine is exploited twice. LDA treatment followed by a borate/hydrogen peroxide introduces the 3-hydroxy substituent. If the pyridine undergoes the 2nd metalation at the 4 position, the alcohol can be protected as a TIPS (triisopropyl silyl) ether, which will force the second metalation to the 5-position. Alternative nitrations may be used, such as converting the lithium intermediate to a stannane and treatment with tetranitromethane, or the use of N02BF| (nitronium tetrafluoroborate) . The Ci displacement may be effected by cuprous cyanide or other sources of cyanide ion. After nitrile hydrolysis and nitro group reduction, ethyl orthoformate may be used instead of -27- formamide for the cyclization, and it may be that some cyclizations will require displacement of F by MeS prior to the reaction. The 4-position is activated by chlorination, and the sidechain amine is then introduced. The final displacement can be by alkoxide or amine nucleophiles to generate the various dialkoxy and amino-alkoxy species, and the appropriate use of R can allow the 7-hydroxyl group to be unmasked at the end of the synthesis. (LDA means lithium diisopropyl amide) . -28- Scheme 4 - Route to Preferred Group 7-See pages 13 and 14.

Use of the 6-alkylquinaldic acid followed by ionic bromination under forcing conditions gives an anhydride, which is opened with ammonia, recyclized to the imide, and then the Hoffman degradation occurs at the less active carbonyl. Cyclization and ring side chain addition in the normal manner is followed by a Stille coupling to introduce the R4 alkyl group. At this step alkenyl or aryl substituents could also be introduced using this coupling technology.

Scheme 5 - Route to Preferred Groups 8, 9, R3 = OR-See page 14.

Dinitration of 2,6-dihydroxypyridine is followed by conversion to the very reactive dichlorocompound. The dinitrodichloropyridine is singly displaced by cuprous cyanide in MP, and then the compound is reduced under mild conditions to the diamine. The nitrile is hydrolysed to the amide, which can then be cyclized to the pyridopyrimidone, which is 4-chlorinated in the usual fashion. Displacement of the more reactive chlorine with the 4-sidechain is followed by displacement of the 6-chlorine with alkoxide. For group 9, the arnine should be alkylated appropriately by methods familiar to one skilled in the art. -29- Scheme 6 - Route to Preferred Group 11-See pages 14 and 15.

Compounds of preferred group 11 are specialized cases of preferred groups 6, 8, 9 and 10, where R3 and R4 are cyclized together. They can be made using the same routes as those described for the preferred groups, with minor modifications, which will be obvious to one skilled in the art. For example vicinally substituted alkoxy amino compounds can be dealkylated, and the corresponding vicinal aminoalcohols can be bisalkylated with an appropriate dihaloalkane .

Scheme 7 - Route for Preferred Groups 12-16-See pages 15 and 16. 2, 4-Diamino-5-cyanopyridine can be cyclized directly to many 4-benzylaminopyridopyrimidine derivatives by treatment with the benzylamine and formic acid at high temperature. For less nucleophilic amines 2, 4-diamino-5-cyanopyridine is converted via ethyl orthoformate/acetic anhydride treatment, followed by cyclization with hydrosulfide ion in anhydrous conditions, to give 7-amino-4-thiono-3H-pyrido [4, 3-d] pyriinidine . S-Alkylation and displacement with an appropriate amine gives the desired product. If R4 is not amino, the amine can be -30- acylated, or reductively alkylated. Alternatively 2 , 4-diainino-5-cyanopyridine can be hydrolysed to the corresponding amide, and this species can be cyclized to 7-amino-4-oxo-3H-pyrido [ , 3-d]pyrimidine with orthoforrnate . Diazotization of the 7-amine and replacement with fluorine allows for introduction of other amine and alkoxide nucleophiles at the end of the synthesis after the C4 substituent has been introduced in the usual manner. Diazotization and replacement of the amine with bromide allows for Stille couplings at the 7-position. -31- Scheme 8 - Route for Preferred Groups 17-21-See pages 16 and 17. 2-Chloro-5-nitropyridine is converted to the corresponding 2-fluorocompound by KF in DMSO. Reduction of the nitro group followed by treatment with Boc anhydride gives the Bocamino derivative, which can be metalated and carboxylated at the 4-position. Removal of the Boc with TFA and cyclization of the pyrimidone ring with formamide gives 6-fluoro-4-oxo-3H-pyrido[3,4-d]pyrimidine. This is 4-chlorinated in the usual manner and the 4-sidechain is introduced via displacement with an appropriate amine. Displacement of the 6-fluorine with appropriate nucleophiles leads to various different final products. If the fluorine is displaced by methanethiol, that in turn can be displaced by alkyl groups in Ni-catalyzed Grignard displacements. -32- Scheme 9 - Route to Preferred Groups 22-26, R4 = H-See pages 17 and 18.

Nitration of 2-methoxynicotinic acid is followed by displacement of the activated xnethoxy group and cyclization of the pyrimidone ring, possibly all in one step with formamidine, or alternatively in two steps with ammonia followed by cyclization with a formamide equivalent. The carbonyl is converted to the chloride and displaced with the sidechain in the usual fashion, and the nitro group is then selectively reduced to amino. This can be alkylated, acylated or diazotized. The diazo compound can be converted to hydroxy or to the bromide or iodide, and these latter can undergo a Stille coupling to introduce lower alkyl, alkenyl, aryl, etc. at R3. -33- Scheme 10 - Route to Preferred Groups 22-26, R3 = H-See pages 17 and 18.

This route uses the known metalation and carboxylation of 2, 6-difluoropyridine, followed by displacement of the 2-fluoro substituent. Cyclization of the pyrimidone ring with formamide, followed by -34- conversion of the carbonyl into chloride in a normal manner gives a chlorofluoropyridopyrirnidine. The ar (alk) ylainino sidechain is introduced by displacement of the more reactive pyrimidine chlorine, and the R4 substituent is then introduced by fluoride displacement. The introduction of alkyl utilizes displacement of F by alkoxide, later ether cleavage to the pyridone, O-triflation and Stille coupling .

Scheme 11 - Route to Preferred Groups 27 and 29-31, R3 = RO-See pages 18 and 19.

This scheme relies on the metalation of 2, 6-difluoropyridine similarly to scheme 10. The first metalation is used to introduce oxygen, and the second to introduce the carboxylic acid. If required to force the second metalation to the 5-position the oxygen may be protected as the very bulky TIPS ether, and stronger bases than LDA may be required. Ammonia is introduced at the 2-position under high temperature and pressure, and the pyridone ring is cyclized, and activated at the 4-position in the usual manner and then displaced with the 4-position sidechain. Displacement of the 7-fluoro substituent with an appropriate nucleophile, followed by conversions as described in previous schemes finishes the synthesis. -35- Scheme 12 - Route to Preferred Group 28-See page 18. 5-Bromo-2,6-difluoronicotinic acid is prepared from 2,6-difluoropyridine by successive lithiations using LDA. The 5-position is alkylated via a Stille coupling, and the pyrimidone ring is cyclized on in two steps. The 4-substituent is introduced in the usual fashion and the 7-fluoro group is displaced with methanethiol. This thioether in turn is displaced by a Grignard agent in the presence of a nickel salt catalyst. Again use of appropriate organometaUic reagents in the Stille and Grignard couplings could lead to alkenyl, alkynyl and aryl substituents at R3 and R4. -36- Scheme 13 - Route to Preferred Groups 29 and 30. R4 = RO-See pages 18 and 19.

Nitration of the commercially available dichloronicotimc acid is followed by a selective displacement of the more reactive CI under mild conditions, followed by a more forcing displacement of the other CI, in the appropriate order. The resulting 6-alkoxy-2-amino-5-nitronicotinic acid is cyclized to the pyrimidone, and the 4-carbonyl is converted to a chloride and displaced in the usual fashion with an appropriate amine to give the 4-amino-7-alkoxy-6-nitropyrido[2,3-d]pyrimidine. Reduction of the nitro group, followed by any desired alkylation or acylation gives the desired compounds.

Scheme 14 - Route to Preferred Group 32-See page 19.

Compounds of group 32 are specialized cases of preferred groups 27, 29, 30 and 31, where R3 and R4 are cyclized together. They can be made using the same routes as those described for these preferred groups with minor modifications. For example, vicinally substituted alkoxy amino compounds can be dealkylated, and the corresponding vicinal amino alcohols can be bisalkylated with an appropriate dihaloalkane. Piperazines can be made by the route shown in Scheme 13, provided that a suitable amine nucleophile is used to displace the 6-chloro substituent instead of an alkoxide. -37- Scheme 15 - Route to Preferred Groups 33-36-See pages 19 and 20.

Reaction of a suitable S-alkylisothiouronium salt with methoxymethylene malononitrile yields a fully functionalized pyrimidine precursor. The initially formed pyrimidine can have the SEt group displaced by R4 either before or after the nitrite hydrolysis, if displacement or oxidation prove problematic later. Displacement of the SEt group can also be achieved without an oxidation to activate the sulfur. Cyclization of the second pyrimidine ring is followed by activation of the 4-carbonyl by thiation and alkylation. Even if the 7-thio group has not been displaced at this point, introduction of the 4-amino side chain occurs preferentially. -38- Scheme 16 - Route to Preferred Groups 37-40-See pages 20 and 21.

The pterine nucleus is made by well-established procedure. For group 37, the pterindione intermediate can be O-alkylated, and for it, and the other groups, the pterindione can be converted to the trichloropterin, and selective displacements can be carried out on the halogens in an order appropriate to give the desired compound.

Scheme 17 - Route to Preferred Groups 41 [3,2-d] ring fusion-See page 21. 3H-Thieno[3/2-d]pyrimid~4-one can be made by standard chemistry from commercially available ethyl 3-aminothiophene carboxylate and formamide. Conversion of the carbonyl to chloride by standard techniques followed by displacement with an appropriate amine gives the desired thieno[3,2-d]pyrimidines. If R4 is not H, an appropriate electrophile, for example nitro for amine based or diazotization derived substituents, or Br for Stille coupled final products, can be introduced either at the stage shown or an earlier stage, and then be converted to R4, by reduction and amination for example or by Stille coupling, or other methods known to those skilled in the art. [This technique follows also for all of the following preferred categories which have the possibility of substitution on R3 or R4, as they all contain electron rich five membered rings which can be readily manipulated by electrophilic aromatic substitution.] (DMSO is dimethyl sulfoxide) . -40- Scheme 18 - Route to Preferred Groups 41 [2, 3-d] ring fusion-See page 21.

Thieno [2, 3-d] pyrimid-4-one is built up by the Ge ald synthesis from 2, 5-dihydroxy-l, 4-dithiane and ethyl cyanoacetate, followed by formarnide cyclization. Conversion of the carbonyl to chloride by standard techniques followed by displacement with an. appropriate amine gives the desired thieno [2, 3-d] pyrimidines .

Scheme 19 - Route to Preferred Groups 42 [3,2-d] ring fusion-See pages 21 and 22.

The [3,2-d] ring fusion compounds are obtained from 3-bromofurfural as shown above in Scheme A. Displacement of the bromide by azide, followed by oxidation of the aldehyde sets up the basic aminofuroic acid needed to fuse on the pyrimidine ring. The annulation shown can be used, or by manipulating which acid derivative is actually used, one could use a variety of other ring annulations, and subsequent activations of the 4-position if required. -41 - Scheme 20 - Route to Preferred Groups 42 [2 -dl ring fusion-See pages 21 and 22.

Reaction of 6-chloro-4-methylthiopyrimidine with LDA followed by DMF gives the corresponding 5-aldehyde,which is treated with the sodium salt of an appropriate glycolate ester, displacing chlorine, and in situ forming the furan ring by intramolecular aldol condensation. Cleavage of the ester and decarboxylation of the unwanted 7-acid functionality may be done in a single reaction with a good nucleophile in a dipolar aprotic solvent at high temperature, or in separate saponification and Cu /quinoline decarboxylation steps. Displacement of the 4-methylthio group by an appropriate amine gives the desired furano[2,3-d]pyrimidines.

Scheme 21 - Route to Preferred Groups 43 [23-dl ring fusion-See page 22.

To make the pyrrolo[2,3-d]pyrimidine a pyrimidine ring is cyclized onto the cyano aminopyrrole using known techniques as shown in the techniques of scheme 2 above. Activation and displacement of the thiol by the side chain can be preceded or followed by the optional electrophilic substitution of the pyrrole ring. -42- Scheme 22 - Route to Preferred Groups 43 [3,2-d] ring fusion-See page 22.

The preparation of the pyrrolo [3, 2-d] pyrimidine exploits the known condensation of orthoforrnate with the acidified 4-methyl group of 6-pyrimidones to form the pyrrolopyrirnidine as shown above. The side chain can be put on by standard techniques such as in Scheme 1, and the R4 substituent can be introduced by standard electrophilic chemistry as described above.

Scheme 23 - Route to Preferred Groups 44 [5,4-dl ring fusion-See page 22.

Condensation of dithioformic acid with 2-aminomalononitrile in the presence of a dehydrating agent such as PPA gives 5-amino-4-cyanothiazole . Reaction of this with orthoforrnate, followed by treatment with MeSNa gives a thiazolo [5, 4-d]pyrimidine derivative, which on treatment with an appropriate amine give the desired compounds . -43- Scheme 24 - Route to Preferred Groups 44 [4,5-d] ring fusion-See page 22.

Reaction of N-cyanobis (methylthio)methyleneimine with ethyl thioglycolate gives ethyl 2-methylthio-4-aminothiazole-5-carboxamide . Cyclization with formamide or equivalent, followed by desulfurization of the methylthio group gives a thiazolopyrimidone, which can be activated by Vilsmeier reagent ,and the chloride displaced by the desired amine to give the desired thiazolo [4 , 5-d]pyrimidine derivatives as shown above.

Scheme 25 - Route to Preferred Groups 45 [5,4-d] ring fusion-See page 22.

The known 5-amino-4-cyanooxazole is treated with ethyl orthoformate/acetic anhydride, and is then reacted with MeSNa to give 4-methylthiooxazolo [5, 4-d] pyrimidine, which on displacement with the appropriate amine gives the desired oxazolo [5, 4-d] pyrimidines as shown above. -44- Scheme 26 - Route to Preferred Groups 45 [4,5-d] ring fusion-See page 22.

Diazotization of the known 5-amino-4, 6-dichloropyrimidine, followed by dilute sulfuric acid treatment give the corresponding 5-hydroxy compound. One of the chlorines is displaced with ammonia, and the oxazole ring is annulated with formic acid or an appropriate equivalent. Displacement of the other chlorine with an appropriate amine gives the desired oxazolo[4, 5-d] pyrimidines as shown above.

Scheme 27 - Route to Preferred Groups 46-See page 23.

These compounds can be made by straightforward displacement of halogen on appropriate 6-chloropurines, by means well documented in the art. R3 substituents can be introduced via facile electrophilic substitutions at the activated 8-position of the purine nucleus, followed by the types of transformation discussed in previous examples.

Scheme 28 - Route to Preferred Groups 47 [5,4-d] ring fusion-See page 23.

Reaction of 6-chloro-4-methylthiopyrimidine with LDA followed by DMF gives the corresponding -45- 5-aldehyde, which is treated with hydroxylamine under mild acidic conditions, and then basic conditions to complete the ring annulation giving 4-Tnethylthioisoxazolo [5, 4-d]pyrirnidine, which on displacement with an appropriate amine gives the desired, isoxazolo [5, -d] pyrimidine derivatives as shown above.

Scheme 29 - Route to Preferred Groups 47 [4, 5-d] ring fusion-See page 23.

Reaction of 4, 6-dichloro-5-nitropyrimidine with CuCN/NMP gives the 4-nitrile. Reduction of the nitro group to the corresponding amine is followed by diazotization and treatment with dilute sulfuric acid to give the corresponding 5-hydroxy compound. Reaction of this with MesAl/ NH4CI gives the amidine which is oxidatively cyclized to 7-amino-4-chloroisoxazolo [4, 5-d] pyrimidine. Removal of the amino f nctionality by diazotization/hypophosphorus acid is followed by displacement of the 4-chlorine with an appropriate amine to give the desired isoxazolo [4 , 5-d] pyrimidine derivatives as shown above. -46- Scheme 30 - Route to Preferred Groups 48 [5, 4-d] ring fusion-See page 23.

Reaction of 6-chloro-4-methylthiopyrimidine with LDA followed by DMF gives the corresponding 5-aldehyde, hich is treated sequentially with. NaSH, NBS and ammonia to complete the ring annulation giving 4-methylthioisothiazolo [5, 4-d] pyrimidine, which on displacement with an appropriate amine gives the desired isothiazolo [5, 4-d] pyrimidine derivatives as shown above.

Scheme 31 - Route to Preferred Groups 48 [4, 5-d] ring fusion-See page 23.

Reaction of 4 , 6-dichloro-5-nitropyrimidine with CuCN/ MP gives the 4-nitrile. Reduction of the nitro group to the amine is followed by diazotization/thiation to give the corresponding 5-mercapto compound. Reaction of this with Me3 l/ NH4CI gives the amidine which is oxidatively cyclized with NBS to 7-amino-4-chloroisothiazolo [4, 5-d] pyrimidine . Removal of the amino functionality by diazotization/hypophosphorus acid is followed by displacement of the 4-chlorine with an appropriate -47- amine to give the desired isothiazolo4, 3-d] pyrimidine derivatives as shown above.

Scheme 32 - Route to Preferred Groups 49 [3, 4-d] ring fusion-See page 23.

Reaction of 6-chloro-4-methylthiopyrimidine with LDA followed by D F gives the corresponding 5-aldehyde, which is treated with hydrazine to do the ring annulation giving 4-inethylthiopyrazolo [3, 4-d] pyriinidine, which on displacement with an appropriate amine gives the desired pyrazolo [3, 4-d] pyrimidine derivatives as shown above.

Scheme 33 - Route to Preferred Groups 49 [4,3-d] ring fusion-See page 23.

Nitration of pyrazole-3-carboxylic acid followed by reduction gives 4-aminopyrazole-3-carboxylic acid. This is cyclized to pyrazolo [ 4 , 3-d] pyrimid-4-one with formamidine HC1, and replacement of the carbonyl with halide by standard procedures, followed by displacement of the chloride by an appropriate amine yields the desired pyrazolo [4 , 3-d] pyrimidine, as shown above. -48- Most Preferred Forms of the Invention 1. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, B, D & E are carbon, A is nitrogen, and R4 is amino. 2. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, B, D & E are carbon, A is nitrogen, and R4 is methylamino. 3. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, B, D & E are carbon, A is nitrogen, and R4 is dimethylamino. 4. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-nitrophenyl, A, D & E are carbon, B is nitrogen, and R4 is amino. 5. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is amino. 6. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 4-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is amino. 7. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is -49- 3-trifluoromethylphenyl, A, D & E are carbon, B is nitrogen, and R4 is amino. 8. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is acetylamino. 9. A most preferred form of the invention is one where X = NH, n = 1 , R1 = H, the aromatic ring is phenyl, A, D & E are carbon, B is nitrogen. 10. A most preferred form of the invention is one where X = NH, n = 1, R1 = H, the aromatic ring is phenyl, A, D & E are carbon, B is nitrogen, and R4 is acetylamino. 1 1. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, B & E are carbon, D is nitrogen, R3 = CI. 12. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, B & E are carbon, D is nitrogen, and R3 is methoxy. 13. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, B & E are carbon, D is nitrogen, and R3 is methylamino. 14. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is -50- 3-bromophenyl, A, B & E are carbon, D is nitrogen, and R3 is dimethylamino.

. A most preferred form of the invention is one where X = H, n = 0, the aromatic ring is 3-bromophenyl, D & E are carbon, and A and B taken together are S. 16. A most preferred form of the invention is one where X = NH, n = 1, R1 =H, the aromatic ring is phenyl, D & E are carbon, and A and B taken together are S. 17. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A & B are carbon, and D and E taken together are S. 18. A most preferred form of the inventio is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, B is carbon, and A, and D and E taken together, are nitrogen. 19. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, B & E are carbon, D is nitrogen, and R3 is N-piperidinyl. 20. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is fluoro. -51- 21. A most preferred form of the invention is one where X = NH , n = 0, the aromatic ring is 3-hydroxyphenyl, A, D & E are carbon, B is nitrogen, and R4 is amino. 22. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is methylamino. 23. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is dimethylamino. 24. A most preferred form, of the invention is one where X = NMe, n = 0, the aromatic ring is 3-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is methylamino. 25. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, D & E are carbon, B is nitrogen, and R4 is methoxy. 26. A most preferred form of the invention is one where X = NH, n = 0, the aromatic ring is 3-bromophenyl, A, B & D are carbon, E is nitrogen, and R4 is fluoro.

Biology These compounds are potent and selective inhibitors of the human EGF receptor tyrosine kinase, -52- and other members of the EGF receptor family, including the Erb-B2, Erb-B3 and Erb-B4 receptor kinases, and are useful for the treatment of proliferative diseases in mammals. These inhibitors prevent mitogenesis in cells where mitogenesis is driven by one or more of this family of receptor kinases. This can include normal cells, where it is desired to prevent mitogenesis, as exemplified by the cells transformed by overexpression or mutation of this kinase family as exemplified by poor prognosis breast cancer where overexpression of EGFR, Erb-B2 and Erb-B3 or mutation of Erb-B2 to the oncoprotein NEU is a major factor in cellular transformation. As the preferred compounds are not highly cytotoxic and do not show potent growth inhibitory properties, because of their high specificity toward inhibition of the EGFR kinase family, they should have a much cleaner toxicity profile than most anti-cancer and anti-proliferative drugs. Their very different mode of action to current anti-cancer drugs should allow for their use in multiple drug therapies, where synergism with available agents is anticipated.

Pharmaceutical compositions and compounds in accordance with the invention have been shown to be very potent, reversible inhibitors of the EGF receptor tyrosine kinase, by binding with high affinity at the adenosine triphosphate (ATP) binding site of the kinase. These compounds exhibit potent IC50S, varying from 10 micromolar to 5 picomolar, for the tyrosine kinase activity of the enzyme, based on an assay examining phosphorylation of a peptide derived from the phosphorylation site of the protein PLCgammal, a known EGFR phosphorylation substrate. This data is shown in Table 1. -53- Biolo2ical Data Materials and Methods Purification of Epidermal Growth Factor Receptor Tyrosine Kinase -Human EGF receptor tyrosine kinase was isolated from A431 human epidermoid carcinoma cells which overexpress EGF receptor by the following methods. Cells were grown in roller bottles in 50% Delbuco's Modified Eagle and 50% HAM F-12 nutrient media (Gibco) containing 10% fetal calf serum. Approximately 109 cells were lysed in two volumes of buffer containing 20 mM 2-(4N-[2-hydroxyethyl]piperazin-l-yl)ethanesulfonic acid (Hepes), pH 7.4, 5 mM ethylene glycol bis(2-aminoethyl ether) Ν,Ν,Ν',Ν'-tetraacetic acid, 1% Triton X-100, 10% glycerol, 0.1 mM sodium orthovanadate, 5 mM sodium fluoride, 4 mM pyrophosphate, 4 mM benzamide, 1 mM dithiothreitol, 80 μg/mL aprotinin, 40 μ ΛηΙ-. leupeptin and 1 mM phenylmethylsulfonyl fluoride. After centrifugation at 25,000 x g for 10 minutes, the supernatant was equilibrated for 2 h at 4°C with 10 mL of wheat germ agglutinin Sepharose that was previously equilibrated with 50 mM Hepes, 10% glycerol, 0.1% Triton X-100 and 150 mM NaCl, pH 7.5, (equilibration buffer). Contaminating proteins were washed from the resin with 1 M NaCl in equilibration buffer, and the enzyme was eluted with 0.5 M N-acetyl-l-D-glucosamine in equilibration buffer, followed by 1 mM urea. The enzyme was eluted with 0.1 mg/ml EGF. The receptor appeared to be homogeneous as assessed by Coomassie blue stained polyacrylamide electrophoretic gels.

WLC 0131 PUS -54- De ermination of ICS0 values - enzyme assays for IC50 determinations were performed in a total volume of 0.1 mL, containing 25 mM Hepes, pH 7.4, 5 mM MgCl2, 2 mM MnCl2, 50 Μ sodium vanadate, 5-10 ng of EGF receptor tyrosine kinase, 200 μΜ of a substrate peptide, (Ac-Lys-His-Lys-Lys-Leu-Ala-Glu-Gly-Ser-Ala-Tyr72 -Glu-Glu-Val- H2-,» derived from the amino acid (Tyr472 has been shown to be one of four tyrosines in PLC (phospholipaseC) -gamma 1 that are phosphorylated by the EGF receptor tyrosine kinase [ ahl, M. I.; Nishibe, S.; Kim, J. W. ; Kim, H. ; Rhee, S. G. ; Carpenter, G., J. Biol. Chem., (1990), 265, 3944-3948.], and peptides derived from the enzyme sequence, surrounding this site are excellent substrates for the enzyme.}, 10 μΜ ATP containing 1 Ci of [32P]ATP and incubated for ten minutes at room temperature. The reaction was terminated by the addition of 2 mL of 75 mM phosphoric acid and passed through a 2.5 cm phosphocellulose filter disc to bind the peptide. The filter was washed* five times with 75 mM phosphoric acid and placed in a vial along with 5 mL of scintillation fluid (Ready gel Beckman) .

WLC 0131 PUS -55- Table 1 EGF Receptor Tyroaine Kinase Inhibition Example # icso 1 8 μΜ ■2.. 3.6 μΜ 3 1.1 μΗ 4 225 nM 5 1.9 μΜ 6 7.6 nM 7 3.1 nM 8 9.6 nM 9 405 nM 10 6.1 μΜ 11 194 nM 12 13 nM 13 250 nM 14 70 nM 15 134 nM 16 3.7 μΜ 17 1.55 μΜ 18 173 nM 19 1.8 /iM 20 4.9μΜ 21 1.25 μΜ 22 39 nM 23 840 nM 24 123 nM 25 377 nM 26 241 nM 27 10 nM WLC 0131 PUS -56- Example # Cso 28 94 nM 29 262 nM 30 10 μΜ 31 15 nM '3-3. 4.7 μΜ 33 130 pM 34 91 pM 35 3.1 nM 36 29 nM 37 39 nM 38 71 nM 39 590 nM 40 578 nM 41 220 nM 42 226 nM 43 10 μΜ ' 44 10 μΜ 45 2.87 μΜ 46 1.42 μΜ 47 1.67 μΜ 48 1.0 μΜ 49 2.5 μΜ 50 10 μΜ 51 1.95 μΜ 52 8 μΜ 53 1.8 μΜ 54 100 ηΜ 55 400 ηΜ 56 110 ηΜ 57 124 ηΜ WLC 0131 PUS -57- Calls Swiss 3T3 mouse fibroblasts, A431 human epidermoid carcinoma cells, and MCF-7 (Michigan Cancer Foundation human mammary carcinoma cells) , SK-BR-3 (human mammary carcinoma cells) , MDA-MB-231 and MDA-MB-468 (human mammary carcinoma cells) breast WLC 0131 PUS -58- carcinomas were obtained from the American Type Culture Collection, Rockville, Maryland and maintained as monolayers in dMEM (Dulbecco's modified eagle medium) /F12, 50:50 (Gibco/BRL) containing 10% fetal bovine serum. To obtain conditioned medium, MDA-MB-231 cells were grown to cofffluency in an 850 cm2 roller bottle and the medium^replaced with 50 ml of serum-free medium. After 3 days the conditioned medium was removed,, frozen down in aliquots and used as a heregulin source to stimulate erbB-2, 3, 4.

Antibodies Monoclonal antibodies raised to the PDGF (platelet-desired growth factor) receptor or phosphotyrosine were from Upstate Biotechnology, Inc., Lake Placid, NY. Anti-pp393ua (antibody to the transcription factor c-jun, which is a 39 kDalton phosphoprotexn) and anti-EGF receptor antibodies were from Oncogene Science, Uniondale, NY.

TTmmiTi^ Tecipitation and Western Blot Cells were grown to 100% confluency in 100 mm Petrie dishes (Coming) . After the cells were treated for 5 minutes with either EGF (epidermal growth factor) , PDGF, or bFGF (basic fibroblast growch factor) (20 ng/ml) or 1 ml of conditioned media from MDA-MB-231 cells, the media was removed and the monolayer scraped into 1 ml of ice cold lysis buffer (50 mM Hepes, pH 7.5, 150 mM NaCl, 10% glycerol, 1% triton X-100, 1 mM EDTA, 1 mM EGTA, 10 mM sodium pyrophosphate, 30 mM p-nitrophenyl phosphate, 1 mM orthovanadate, 50 mM sodium fluoride, 1 mM WLC 0131 PUS -59- phenylmethylsulfonylfluoride, 10 μg/ml of aprotinin, and 10 μ /ml of leupeptin) . The lysate was transferred to a microfuge tube (small centrifuge that holds 1-2 ml plastic centrifuge tubes) , allowed to sit on ice 15 minutes and centrifuged 5 minutes at 10,000 x g. The supernatant was transferred to a clean microfuge tube and 5 g of antibody was added to designated samples. The tubes were rotated for 2 hours at 4° C after which 25 μΐ of protein A sepharose was added and then rotation continued for at least 2 more hours. The protein A separose was washed 5 times with 50 mM Hepes, pH 7.5, 150 mM NaCl, 10% glycerol and 0.02% sodium azide. The precipitates were resuspended with 30 μΐ of Laemlli buffer (Laemmli, NATURE, Vol. 727, pp. 680-685, 1970), heated to 100'C for 5 minutes and centrifuged to obtain the supernatant. Whole cell extracts were made by scraping cells grown in the wells of 6 well plates into 0.2 ml of boiling Laemmli buffer. The extract were transferred to a microfuge tube and heated to 100° C for 5 minutes. The entire supernatant from 'the immunoprecipitation or 35 μΐ of the whole cell extract was loaded onto a polyacrylamide gel (4-20%) and electrophoresis carried out by the method of Laemlli (Laemmli, 1970) . Proteins in the gel were electrophoretically transferred to nitrocellulose and the membrane was washed once in 10 mM Tris buffer, pH 7.2, 150 mM NaCl, 0.01% Azide (TNA) and blocked overnight in TNA containing 5% bovine serum albumin and 1% ovalbumin (blocking buffer) . The membrane was blotted for 2 hours with the primary antibody (^g/ml in blocking buffer) and then washed 2- times sequentially in TNA, TNA containing 0.05% Tween- 20 and 0.05% Nonidet P-40 (commercially available detergent) and TNA. The membranes were then incubated for 2 hours in blocking buffer containing 0.1 /xCi/ml of [125I] protein A and then washed again as above.

WLC 0131 PUS -60- After the blots were dry they were loaded into a film cassette and exposed to X-AR X-ray film for 1-7 days. Protein A is a bacterial protein that specifically bonds certain IgG subtypes and is useful in binding to and isolating antibody-antigen complexes.

Northern Biota Total cellular RNA was isolated from untreated control or treated Swiss 3T3 cells using RNAzol-B (trademark of Tel Test Inc. for a kit used to isolate RNA from tissues) and adhered to the protocol described by the manufacturer. Forty to fifty ^g of RNA was loaded onto a 1% agarose gel and electrophoresis carried out for 3-4 hours at 65 volts. The RNA in the gel was transferred by capillary action to a nylon membrane (Hybond-N, Amersham) . The 40 mer c-jun probe was end labeled with [32P]ATP using T4 nucleotide kinase (Promega) and purified on a G25 sephadex column according to the procedure recommended by the supplier, Oncogene Science. Hybridization was performed overnight at 65 'C (c-jun is an immediate early transcription factor; it is one of the components of AP-1 while FOS is the second component of AP-1.

Growth Factor-Mediated Mitoaenesis Swiss 3T3 fibroblasts were grown to 90 - 100% confluency in 24- well plates (1.7 x 1.6 cm, flat bottom) and growth arrested in serum-free media for 18 hours. Drug was added to specified wells 2 hours prior to growth factors and then the cells were exposed to either 20 ng/ml EGF, PDGF or bFGF or 10% serum for 24 LC 0131 PUS -61- hours. Two ^Ci of [methyl-3H] thymidine was added to each well and incubated for 2 hours at 37° C. The cells were trypsinized and injected into 2 ml of ice cold 15% trichloroacetic acid (TCA) . The resulting precipitate was collected on glassfiber filters, washed five times with 2-ml aliquots of ice-cold 15% TCA, dried and placed in scintillation vials along with 10 ml Ready gel (Beckman, Irvine, CA) .

Radioactivity was determined in a Beckman LS 6800 scintillation counter.

Growth Inhibition Assay Cells (2 x 104) were seeded in 24-well plates (1.7 x 1.6 cm, flat bottom) in two mis of medium with or without various concentrations of drug. Plates were incubated for 3 days at 37o in a humidified atmosphere containing 5% C02 in air. Cell growth was determined by cell count with a Coulter Model AM electronic cell counter (Coulter Electronics, Inc., Hialeah, FL) .

INHIBITION OP EGF-INDUCED AUTOPHOSPHORYLATION IN A431 EPIDERMOID CARCINOMA CELLS AND CONDITIONED MEDIA-INDUCED AUTOPHOSPHORYLATION IN SK-BR-3 BREAST TUMOR CELLS BY COMPOUNDS OP THE CURRENT INVENTION Example # EGFR ICS0 nM A431 ICS0 nM SKBR-3 ICS0 nM 4 225 >1000 >10, 000 6 7.6 53 2660 7 3.1 20 100 8 9.6 32 71 22 39 252 -1500 27 10 110 -800 59 2.6 12 <10 -62- ANTIPROLIFERATIVE PROPERTIES OF TYROSINE KINASE INHIBITORS IC50 (nM) B104-1-1 - NIH-3T3 fibroblasts transfected by the neu oncogene, Stem et al., Science, 234, pp. 321-324 (1987) SK-BR-3 - Human breast carcinoma overexpressing erbB-2 and erbB-3 MDA-468 - Human breast carcinoma overexpressing the EGF receptor The above gels, developed as detailed in the experimental section, demonstrate the efficacy of compounds used in the pharmaceutical compositions of the current invention at blocking certain EGF-stimulated mitogenic signalling events in whole cells. The numbers to the left of the gels indicated the positions of molecular weight standards in kiloDaltons. The lane labelled control shows the degree of expression of the growth-related signal in the absence of EGF stimulation, whereas the lane labelled EGF (or PDGF or b-FGF) shows the magnitude of the growth factor-stimulated signal. The other lanes show the effect of the stated quantities of the named drug on the growth factor-stimulated activity being -63- measured, demonstrating that the compounds used in the pharmaceutical compositions of the present invention have potent effects in whole cells, consistent with their ability to inhibit the tyrosine kinase activity of the EGF receptor.

Gel of Example 40 (Fig. 7) detects mRNA for c-jun by hybridization with a specific radiolabelled RNA probe for c-jun. The gel demonstrates that the growth factors EFG, PDGF and b-FGF stimulate c-jun production in Swiss 3T3 cells, and that compound 40 blocks this production for EGF-stimulated cells, but not for PDGF or b-FGF stimulated cells.

Effect of Example 40 on Growth Factor Mediated Expression of p39c Jun This gel shows the amount of c-jun induced in Swiss 3T3 cells by the growth factor EGF, PDGF and b-FGF, quantitating with an anti-c-jun-specific monoclonal antibody. It demonstrates the ability of Example 40 to block c-jun expression in Swiss 3T3 when stimulated by EGF, but not when stimulated by PDGF or b-FGF.

It is to be appreciated that the compounds described herein can be used in combination with other components to enhance their activity. Such additional components are anti-neoplastic materials as, doxorubicin, taxol, cis platin, and the like.

It has been found that the compounds described herein may inhibit both the erb-B2 and erb-B4 receptors and therefore have significantly increased clinical activity advantageously in WLC 0131 PUS -64- combination with the aforementioned anti-neoplastic agents .

See also the results shown in Figures 1 - 17.

Some preferred structures are as follows: Ex.# Z 4 - fluorine 6 - NH2 7 - HCHj 8 - N (CH3)2 WLC 0131 PUS -65- Ex # Z 22 H2 -N02 27 NH, Br Ex # Z 59 - OCH3 Br 60 - NH CH3 Br 61 - N(CH3)2 Br Chemical Experimental Listed below are preferred embodiments wherein all temperatures are in degrees Centigrade all parts are parts by weight unless otherwise indicated.

WLC 0131 PUS -66- Example 1 4 -Anilinopyrido Γ3 , 2 -dl pyrimidine mesylate 3H-Pyrido f3 , 2-dl pyrimidin-4 -one . A solution of 6-chloro-3-nitropicolinamide (2.00 g, 9.91 mmol) in EtOAc/MeOH (1:1, 100 mL) is hydrogenated over 5% Pd-C (0.40 g) at 60 psi for 6 days, with additions of fresh catalyst after 2 and 4 days . After removal of the catalyst by filtration the solution is concentrated to dryness, to give 3 -aminopicolinamide as an orange oil, which is used directly in the next step. The crude product is stirred under reflux with triethyl orthoformate (50 mL) for 42 h, during which time a tan precipitate forms. After cooling, the solid is filtered off, washed well with petroleum ether, and dried under vacuum to give 3H-pyrido [3 , 2-d] pyrimidin-4-one (1.27g, 87%), mp 343-345 °C [Price, C.C. and Curtin, D.Y. J. Amer. Chem. Soc. 68, 914, 1946 report mp 346-347 °C] . 4-ChloroPyrido Γ3.2-dl pyrimidine . A suspension of the above pyrimidinone (1.00 g, 6.80 mmol) in Ρ0(ΙΓ3 (30 mL) is heated under reflux for 4 h, and then concentrated to dryness under reduced pressure. The residue is partitioned between CH2Cl2 and saturated NaHC03. solution, and the organic layer worked up to give 4 -chloropyrido [3 , 2-d] pyrimidine (0.97 g, 86%) as a tan solid, mp 335 °C (dec) , which is used without further characterisation. 4 -Anilinopyrido Γ3 , 2-dl pyrimidine mesylate . A solution of 4 -chloropyrido [3 , 2-d] pyrimidine (84 mg, 0.5 mmol), aniline (56 mg, 0.6 mmol) and triethylamine (62 mg, 0.6 mmol) in EtOH (2 mL) are refluxed under N2 WLC 0131 PUS -67- with stirring for 2 h. The crude reaction mixture is purified on a preparative tic plate (silica) , eluting once with 3% MeOH in CHC13. The major band is extracted, and evaporated to dryness under reduced pressure, and the residual solid is dissolved in acetone, (5 mL) , filtered, and methanesulfonic acid (32 μΐι, 0.5 mmol) is added slowly with swirling. The precipitate is collected by suction filtration, rinsed with acetone and dried in a vacuum oven to give 4-anilinopyrido [3 , 2-d] pyrimidine mesylate (91 mg, 57%) as dull yellow needles. rH MR (D SO) δ 11.75 (1H, slbrs) , 9.11 (1H, dd, J = 1.5, 4.3 Hz), 8.97 (1H, s) , 8.32 (1H, dd, J = 1.5, 8.4 Hz), 8.12 (1H, dd, J = 4.3, 8.5 Hz), 7.88 (2H, d, J = 8.2 Hz) , 7.49 (2H, t, J = ' 8.0 Hz), 7.32 (1H, t, J = 7.0 Hz) , 2/34 (3H, s) .

Example 2 4 -Benzylaminopyrido [3 , 2 -dl pyrimidine A solution of freshly prepared 4- chloropyrido [3 , 2-d] pyrimidine (0.10 g, 0.60 mmol) (prepared as described in the previous experimental) and benzylamine (0.13 mL, 1.20 mmol) in propan-2-ol (15 mL) containing a trace of cone. HC1 is warmed at 50 °C for 30 min, and then concentrated to dryness. The residue is partitioned between water and EtOAc, and the organic layer worked up and chrpmatographed on silica gel. EtOAc elutes foreruns, while MeOH/EtOAc (1:9) elutes 4- (benzylamino) pyrido [3 , 2-d] pyrimidine (0.11 g, 77%). *H NMR (CDC13) δ 8.67 (1H, s) , 6.50 (1H, dd, J = 4.3, 1.5 Hz), 8.10 (1H, dd, J = 8.5, 1.5 Hz), 7.63 (1H, dd, J = 8.8, 4.3 Hz), 7.55 (1H, brs) , 7.41-7.29 (5H, m) , 4.86 (2H, d, J = 5.9 Hz).

WLC 0131 PUS -68- Example 3 4- (3-Bromoanilino) pyrido f3.2 -dl pyrimidine Reaction of 4-chloropyrido [3 , 2-d] yrimidine (prepared as described in a previous experimental) with 3-bromoaniline in propan-2-ol containing a trace of cone. HC1 at 50 °C for 30 min, followed by chromatography of the product on silica gel, gives 4- (3 -bromophenyl) aminopyrido [3 , 2-d] pyrimidine (87% yield). XH NMR (CDC13) δ 9.19 (1H, brs) , 8.83 (1H, s) , 8.80 (1H, dd, J = 4.3, 1.5 Hz), 8.29 (1H, brs), 8.19 (1H, dd, J = 8.5, 1.5 Hz), 7.83 (1H, m) , 7.76 (1H, dd, J = 8.5, 4.3 Hz), 7.29-7.27 (2H, m) .

Example 4 4- (3-Bromoanilino) -6-fluoropyrido ("3 , 2-dl pyrimidine 2 -cvano-6-fluoro-3 - itropyridine . A mixture of 6-chloro-2-cyano-3-nitropyridine [Colbry, N. L.; Elslager, E. F. ; erbel, L. M. ,· J. Het. Che . , 1984, 21, 1521-1525] (10 '0 g, 0.054 mol) and KF (9.48 g, 0.163 mol) in MeCN (200 mL) is heated under reflux with stirring, for 18 h, then poured into water and extracted with EtOAc. The extract is washed with water and worked up, and the residue is chromatographed on silica gel, eluting with EtOAc/petroleum ether (3:7), to give after removal of the solvent under reduced pressure 2-cyano-6-fluoro-3-nitropyridine (7.2 g, 79%) . :H NMR (CDClj) δ 8.79 (1H, dd, J = 9.0, 6.0 Hz) , 7.48 (1H, dd, J = 9.0, 3.0 Hz). 6-Fluoro-3-nitropyridine-2 -carboxamide . A solution of 2-cyano-6-fluoro-3 -nitropyridine (1.40 g. 8.39 mmol) in 90% H2S04 (30 mL) is warmed at 70 °C for WLC 0131 PUS -69- 90 min, then cooled, poured onto ice and basified with cone, ammonia. Extraction with EtOAc and workup gives 6-fluoro-3-nitropyridine-2-carboxamide (0.94 g, 61%) . XH MR (CDClj) 6 8.70 (1H, dd, J = 8.9, 6.5 Hz), 8.30, 8.03 (1H, 1H, brs) , 7.62 (1H, dd, J = 8.9, 2.9 Hz). 6-Fluoro-3H-pyrido Γ3.2-dl pyrimid-4-one . A solution of 6-fluoro-3 -nitropyridine-2-carboxamide (1.50 g, 8.10 mmol) in EtOAc (80 mL) is hydrogenated over 5% Pd-C (0.30 g) at 60 psi for 2 h. After removal of the catalyst by filtration, the solvent is removed under reduced pressure, to give a residue of crude 3-amino-6-fluoropyridine-2-carboxamide which is used directly in the next step. Triethyl orthoformate (60 mL) is added and the mixture is then heated under reflux with vigorous stirring for 18 h. The cooled mixture is diluted with an equal volume of petroleum ether, and the resulting precipitate collected by filtration and is washed well with petroleum ether to give 6-fluoro-3H-pyrido [3 , 2-d] pyrimid-4-one (1.26 g, 84%). Ή NMR (DMSC¾ δ 12.72 (1H, brs) , 8.31 (1H, dd, J = 8.6, 7.7 Hz), 8.20 (1H, s) , 7.66 (1H, dd, J = 8.6, 3.0 Hz) . 4 - ( 3 -Bromoani1ino) -6-fluoropyrido f3.2 -d) pyrimidine . A suspension of 6-fluoro-3H-py ido [3 , 2-d] pyrimid-4-one (0.20 g, 1.21 mmol) in P0C13 (30 mL) is heated under reflux with stirring until homogeneous (2 h) , and then for a further 1 h. Excess POCl3 is removed under reduced pressure, and the residue is partitioned between CH2C12 and saturated aqueous NaHC03. Workup of the organic portion gives crude 4-chloro-6- fluoropyrido [3 , 2-d] pyrimidine (100%) as an unstable white solid which is used directly in the next step.

WLC 0131 PUS -70- A solution of 4-chloro-6-fluoropyrido [3 , 2-d]pyrimidine (0.20 g, l.l mmol) and 3 -bromoaniline (0.12 mL, 2.18 mmol) in propan-2-ol (20 mL) containing cone. HC1 (l drop) is heated under reflux for 15 in, then cooled, poured into water and extracted with EtOAc. The extract is worked up, and the residue chromatographed on silica gel, eluting with EtOAc/petroleum ether (1:2) to give after removal of the solvent under reduced pressure 4- (3 -bromoanilino) -6 -fluoropyrido [3, 2-d) yrimidine (0.18 g, 52%). XH NMR (CDC13) δ 8.82 (1H, s) , 8.65 (1H, brs) , 8.31 (1H, t, J = 7.4 Hz), 8.27 (1H, brs), 7.77 (1H, m) 7.41 (1H, dd, J = 8.9, 2.2 Hz), 7.29 (2H, brs).

Example 5 4- (3 -Bromoanilino) - 6 -chloropyrido Γ3.2 -dl pyrimidine 6 -chloro- -nitropicolinamide . A solution of 6-chloro-3 -nitropicolinonitrile (1.00 g, 5.45 mmol) in 90% H2S04 (15 mL) is warmed at 70 °C for 3.5 h, and then poured into .;ce-water. The mixture is extracted four times with EtOAc and the combined extracts worked up to give 6 -,chloro-3 -nitropicolinamide (0.80 g, 73%) . XH NMR (DMSO) δ 8.55 (1H, d, J = 8.5 Hz), 8.31, 8.04 (1H, 1H, 2 brs), 7.93 (1H, d, J = 8.5 Hz). 6-Chloro-3H-pyrido f3 , 2-d) pyrimidin-4-one . A solution of 6-chloro-3 -nitropicolinamide (0.30 g, 1.49 mmol) in EtOAc (30 mL) is hydrogenated at 60 psi over 5% Pd-C (0.10 g) for 20 min. After removal of the catalyst by filtration the solution is concentrated to dryness to give 3 -amino- 6 -chloropicolinamide as a yellow oil, which is used directly in the next step. It is dissolved in triethylorthoformate (30 mL) and LC 0131 PUS -71- the mixture is heated under reflux for 18 h.

Petroleum ether (30 mL) is added to the cooled solution, and the resulting precipitate of crude 6-chloro-3H-pyrido [3 , 2-d) pyrimidin-4-one (0.27 g, 99%) is filtered off and dried in a vacuum oven. 4 - ( 3 -Bromoani1ino) -6 -chloropyrido f3.2 -dl pyrimidine . A suspension of the above quinazolone (0.20 g, 1.10 mmol) in P0C13 (30 mL) is heated under reflux for 3 h, and then concentrated to dryness under reduced pressure. The residue is partitioned between CH2C12 and saturated NaHC03 solution, and the organic portion is worked up to give 4 , 6-dichloropyrido [3 , 2-d] pyrimidine (0.16 g, 73%) as a tan solid, which is used directly in the next step. A solution of the crude dichloropyridopyrimidine (0.16 g, 0.80 mmol) and 3-bromoaniline (0.17 mL, 1.60 mmol) in propan-2-ol (25 mL) containing a trace of cone. HC1 is warmed at 50 °C for 30 min. The cooled mixture is poured into saturated NaHC03 and extracted with EtOAc, and the extract is worked 'up and chromatographed on silica gel. Elution with EtOAc/petroleum ether (1:4) gives 3-bromoaniline, while EtOAc/petroleum ether (1:1) elutes 4- (3 -bromoanilino) -6-chloropyrido [3 , 2-dlpyrimidine (0.17 g, 63%). . Ή NMR (CDC13) δ 8.90 (1H, brs,) 8.84 (1H, s) , 8.30 (1H, dd, J = 2.1, 2.0 Hz) 8.17 (1H, d, J = 8.8 Hz), 7.82-7.78 (1H, m) 7.73 (1H, d, J = 8.8 Hz), 7.32-7.29, (2H, m) .

Example 6 4- (3-Bromoanilino) -6-aminopyrido Γ3.2-dl pyrimidine Reaction of 4- (3-bromoanilino) -6-fluoropyrido [3 , 2-d) pyrimidine (0.12 g, 0.38 WLC 0131 PUS -72- mmol) (described in a previous experimental) with a saturated solution of ammonia in ethanol in a pressure vessel at 100 °C for 18 h gives 6-amino-4- (3-bromoanilino) pyrido [3, 2-d] pyrimidine, (87 mg, 72%) . 1H MR (CDC13) δ 8.76 (1H, brs) , 8.64 (1H, s) , 8.23 (1H, brs) , 7.93 (1H, d, J « 9.0 Hz), 7.81 (1H, dt, Jd = 7.7 Hz, Jt =1.8 Hz), 7.28-7.22 (2H, m) , 7.00 (1H, d, J = 9.0 Hz) , 4.90 (2H, brs) .

Example 7 4- (3 -Bromoanilino) -6-methylaminopYrido [3.2-dl pyrimidine Reaction of 4- (3-bromoanilino) -6-fluoropyrido [3 , 2-d) pyrimidine (50 mg, 0.16 mmol) (described in a previous experimental) with methylamine hydrochloride (32 mg, 0.47 mmol) and triethylamine (70 μΐι, 0.55 mmol) in ethanol (10 mL) in a pressure vessel at 100 °C for 18 h gives 6-methylamino-4- (3-bromoanilino) pyrido [3 , 2-d] pyrimidine (43 mg, 81%). XH 'NMR (CDC13) δ 8.81 (1H, brs), 8.61 (1H, s) , 8.19 (1H, t, J = 1.8 Hz), 7.86 (1H, d, J = 9.1 Hz,), 7.¾3 (1H, dt, Jd = 7.7 Hz, Jc =1.8 Hz), 7.28-7.21 (2H, m) , 6.92 (1H, d, J = 9.1 Hz), 4.97 (1H, q, J = 5.0 Hz) , 3.13 (3H, d, J = 5.0 Hz) .

Example 8 4- (3 -Bromoanilino) -6-dimethylaminoPyrido Γ3.2-dl pyrimidine .

A mixture of 4 - (3 -bromoanilino) -6- fluoropyrido [3 , 2-d) pyrimidine (0.15 g, 0.47 mmol) (described in a previous experimental) , dimethylamine hydrochloride (0.11 g, 1.41 mmol) and triethylamine WLC 0131 PUS -73- (0.23 mL, 1.64 mmol) in EtOH (15 mL) is heated in a pressure vessel at 100 °C for 18 h. The solvent is removed under reduced pressure, and the residue is partitioned between EtOAc and water. The organic portion is worked up, and the residue chromatographed on silica gel. Elution with EtOAc/petroleum ether (1:1) gives foreruns, while EtOAc elutes off 4- (3-bromoanilino) -6-dimethylaminopyrido [3 , 2-d] yrimidine (0.14 g, 86%). XH NMR (CDC13) δ 8.72 (1H, brs) , 8.56 (1H, s) , 8.17 (1H, t, J = 1.9 Hz) , 7.85 (1H, d, J" = 9.3 Hz), 7.77 (1H, dt , Jd = 7.5 Hz, Jt =1.9 Hz), 7.27-7.18 (2H, m) , 7.08 (1H, d, J = 9.3 Hz), 3.21 (6H, s) .

Example 9 4- (3-Bromoanilino) -6 -methoxypyrido Γ3.2 -dl pyrimidine 4- (3-Bromoanilino) -6-fluoropyrido [3,2-d] pyrimidine (described in a previous experimental) (0.11 g, 0.34 mmol) is added to a solution of NaOMe (prepared by the addition of Na metal (31 mg, 1.38 mmol) to dry eOH*(15 mL) . After heating in a pressure vessel at 90 °C for 3 h, the solution is concentrated to dryness and the residue is partitioned between EtOAc and water. Workup of the organic portion gives 4 - ( 3 -bromophenyl ) amino- 6 -methoxypyrido [3,2-d]pyrimidine (92 mg, 82%). XH NMR (CDC13) δ 8.73 (1H, s), 8.66 (1H, brs), 8.18 (1H, m) , 8.05 (1H, d, J = 8.9 Hz), 7.83-7.80 (1H, m) , 7.30-7.24 (2H, m) , 7.23 (1H, d, J = 8.9 Hz) , 4.12 (3H, s) .

WLC 0131 PUS -74- Example 10 4 -Anilinopyrido ί4 , 3 -dl pyrimidine 4 - (N-1-Butoxycarbonylamino) yridine . To a mixture of 4-aminopyridine (2 g, 21.24 mmol) , potassium hydroxide (3.57 g, 63.72 mmol), water (10 mL) , and 2-methyl-2-propanol (4 mL) on ice is added di-t-butyl-dicarbonate (6.95 g, 31.87 mmol). The resulting biphasic solution is stirred at 25 °C for 1 week, then water (20 mL) is added. The solution is extracted with IX CH2C12 and 2X EtOAc. The organic layer is dried (MgS04) and concentrated under reduced pressure to give 4- (N-t-butoxycarbonylamino) pyridine (4.08 g, 99%). 1H NMR (DMSO) δ 9.84 (1H, s) , 8.35 (2Η,' d, J = 6 Hz) , 7.44 (2H, d, J = 7 Hz) , 1.49 (9H, s) . 4- (N- -Butoxycarbonylamino) nicotinic acid. n-Butyl lithium (2.18 M, 24 mL, 52.51 mmol) is added slowly to a solution of 4- (N-t-butoxycarbonylamino) -pyridine (4.08 g, 21 mmol) in THF (50 mL, stirred under N2 at -78 'c The solution is allowed to warm to 0*C, stirred for 3 h, then cooled again to -78 *C and poured into e her (100 mL) containing dry ice. The solution is warmed to room temperature with constant stirring. Water is added and the mixture is neutralized with acetic acid. The resulting solid is col-lected by vacuum filtration and dried in a vacuum oven to give 4- (N-t-butoxycarbonylamino) nicotinic acid (2.72 g, 54%) as a brown solid. XH NMR (DMSO) δ 11.75 (1H, brs) , 8.95 (1H, s) , 8.50 (1H, d, J = 6.0 Hz), 8.20 (1H, d, J = 6.0 Hz), 1.49 (9H, s) . 4 -Amino nicotinic acid. A mixture of 4- (N-t-butoxycarbonylamino) nicotinic acid (2.72 g, 11.4 -75- mmol), TFA (10 mL), and CH2C12 (20 mL) is stirred at room temperature for 12 h. The volatiles are removed under reduced pressure, and the resulting crude 4-amino nicotinic acid is used directly in the next reaction. 3H-Pyridoi4,3-d pyrimidin-4-one. Crude 4-amino nicotinic acid (2.72 g, 11.4 mmol) in formamide (20 mL) is heated to 170°C for 12 h. The volatiles are distilled out under reduced pressure (0.8 mmHg). The residual solid is then purified on a medium pressure silica gel column, eluting with 10% MeOH in CHC13 to give 3H-pyrido[4,3-d]pyrimidin-4-one (780 mg, 47%) as a whitish yellow solid. 1H M (DMSO) 5 12 . 64 , (1H, brs), 9.28 (1H, s), 8.83 (1H, d, J = 5.5 Hz), 8.30 (1H, s), 7.58 (lH, d, J = 5.8 Hz). 3H-Pyridor4,3-dlpyrimidine-4-thione. Phosphorus pentasulfide (2.59 g, 5.83 mmol) is added to a solution of 3H-pyrido[4,3-d]pyrimidin-4-one (780 mg, 5.3 mmol) in pyridine (5 mL). The mixture is refluxed for 5 h. On cooling a precipitate forms and the supernatant is decanted off. The solid is suspended in water (20 mL) and then filtered to yield 3H-pyrido[4,3-d]pyrimidine-4-thione (676 mg, 78%) as a black solid. 1H NMR(DMSO) δ 14 . 53, (1H, brs), 9.65 (1H, s), 8.84 (1H, d, J = 7.0 Hz), 8.32 (1H, s), 7.64 (1H, d, J = 8.0 Hz) . 4-Methylthiopyrido 4.3-d]pyrimidine. A mixture of 3H-pyrido[4,3-d]pyrimidine-4-thione (676 mg, 4.14 mmol), triethylamine (1.4 mL, 10.31 mmol), DMSO (4 mL), and iodomethane (0.48 mL, 7.72 mmol) is stirred for 12 h under N2 at 25°C. The mixture is WLC 0131 PUS -76- poured onto water and extracted with EtOAc. The organic extracts are dried (MgS04) , and the solvent is removed under reduced pressure to yield 4-methylthiopyrido [ , 3-d] yrimidine (1.15 g# quant.) as a brown solid. XH MR (DMSO) 6 9.52 (1H, s) , 9.16 (1H, s) , 8.95 (1H, d, J = 6 Hz) , 7.86 (1H, d, J = 8 Hz) , 2.75 (1H, s) . 4-Anilinopyrido [4.3-d] pyrimidine . A mixture of 4 -methylthiopyrido [4 , 3 -d] pyrimidine (174 mg, 0.97 mmol) , and aniline (186.2 mg, 1.99 mmol) in EtOH (2 mL) is refluxed under N2 for 12 h. Cooling to 0*C forms a solid which is filtered to yield 4-anilinopyrido- [4, 3-d] yrimidine (34.5 mg, 16%). ¾ NMR (DMSO) δ 10.29 (1H, brs) , 9.86 (1H, s) , 8.82 (1H, d, J = 5.8 Hz), 8.72 (1H, s) , 7.85 (2H, d, J = 7.5 Hz) , 7.66 (1H, d, J = 5.5 Hz), 7.45 (2H, t, J = 8.0 Hz), 7.23 (1H, t, J = 7.3 Hz) .

Example 11 4- (3-Bromoanilino'pyrido [4.3-dl pyrimidine A mixture of 4 -methylthiopyrido [4 , 3-d]pyrimidine (171 mg, 0.96 mmol), (see previous experimental) and 3 -bromoaniline (1 mL) is heated to 100 *C for 2 h. A solid precipitates on cooling and is collected by vacuum filtration and then recrystallized from EtOH to yield 4 - (3 -bromoanilino) pyrido [4 , 3 -d]pyrimidine (30 mg, 10%). XH NMR (DMSO) δ 10.33 (1H, s) , 9.86 (1H, s) , 8.84 (1H, d, J = 5.8 Hz), 8.79 (1H, S) , 8.22 (1H, s) , 7.89 (1H, d, J = 7.2 Hz) , 7.69 (1H, d, J = 5.8 Hz), 7.40 (2H, dt, Jd = 8.0 Hz, Jt = 1.5 Hz) . -77- Example 12 4- ( 3-Bromoanilino) -7-fluoropyrido [4 , 3-d] yrimidine 3-Cyano-4, 6-diami.nopyridine . Crude 2-bromo-3-cyano-4 , 6-diaminopyridine [W. J.Middleton, US Patent 2,790,806 (April 30, 1957), Du Pont; Chem. Abst. 51:P14829 (1957), see also next experimental] (15.1 g, 0.071 mole) is hydrogenated in THF/MeOH (200 mL, 2:1) containing KOAc (7.0 g, 0.071 mole) and 5% Pd/C (4 g) at 55 p.s.i. and 20°C for 7 days. Filtration over Celite, washing with THF/MeOH and removal of the solvent gives a solid, which is dissolved in dilute HC1 and water. Adjustment of the solution pH to 10 (cone. NaOH) and cooling gives 3-cyano-4 , 6-diaminopyridine (6.58 g, 69%) as a yellow solid, mp 197-198°C [Metzger, R. Oberdorfer, J.; Schwager, C; Thielecke, W.; Boldt, P. Liebigs Ann. Chem. 1980, 946-953 record mp (benzene) 205°C] . Extraction of the remaining liquor with EtOAc (4 x 200 mL) gives further product (2.12 g, 22%). XH NMR (DMSO) δ 7.91, ( 1H, s) , 6.26, 6.24 (2H, 2H, brs), 5.63 (1H, s) . 4 , 6-Diamino-3-pyridylcarboxamide . 3-Cyano- , 6-diaminopyridine (4.30 g, 0.032 mole) is added to 90% H2S04 (25 mL) , then stirred at 60-70°C for 3 h. The resulting solution is added to cold cone. NaOH (40%) to give a mixture of , 6-diamino-3-pyridylcarboxamide and inorganic salts. An analytically pure sample is obtained by chromatography on alumina (10-50% MeOH/CHCl3) to give a pale yellow solid. XH NMR (DMSO) δ 8.15, (lH,s), 6.91 (2H, brs), 7.7-6.3 (2H, brm) , 5.78 (2H ,brs), 5.56 (1H, s).

WLC 0131 PUS -78- 7-Amino-4-oxo-3tf-Dyrido Γ4.3-dl pyrimidine . Crude 4 , 6-diamino-3-pyridylcarboxamide (9.2 g) is heated in purified (EtO)3CH (distilled from Na, 60 mL) at 170 °C for 1.5 d. After removing the solvent, the residue is dissolved in hot 2 M NaOH, filtered, neutralized (cone. HCl) and cooled to give 7-amino-4-oxo-3H-pyrido [4, 3-d] pyrimidine (3.57 g, 69% from the nitrile) as a light brown solid ¾ NMR (DMSO) δ 11.79 (1H, brs) , 8.74 (1H, s) , 7.97 (1H, s) , 6.76 (2H, brs) , 6.38 (1H, s) . 7-Fluoro-4-oxo-3tf-pyrido Γ4.3-dl pyrimidine . A solution of 7-amino-4-oxo-3H-pyrido [4, 3-d] yrimidine (1.00 g, 6.17 mmol) in 60% HBF4 (25 mL) at 0 °C is treated with solid NaN02 (0.85 g, 12.3 mmol, added in portions over 2 h) , and is then stirred at 0 °C for a further 1 h and at 20 °C for 30 min. The resulting mixture is ice-cooled, neutralized with saturated aqueous Na2C03, and extracted with EtOAc (4 x 100 mL) . The extract is washed with water, then filtered through silica gel* (EtOAc) to give 7-fluoro-4-oxo-3H-pyrido [4, 3-d] yrimidine (0.48 g, 47%) as a cream solid. XH NMR* (DMSO) 6 12.69 (1H, brs) , 9.01 (1H, s) , 8.31 (1H, s) , 7.34 (1H, s) 4- (3-Bromoanilino) -7-fluoropyrido Γ4.3-dl pyrimidine . A suspension of 7-fluoro-4-oxo-3H-pyrido [4 , 3-d] pyrimidine (0.23 g, 1.39 mmol) in POCl3 (10 mL) is stirred under reflux for 3.5 h, and is then concentrated under vacuum. The resulting oil is ice-cooled, diluted with CH2C12 (100 mL) , saturated aqueous Na2C03 (40 mL) and ice, and stirred at 20 °C for 2 h.

The CH2C12 extract is separated and the aqueous portion further extracted with CH2C12 (2 x 100 mL) , and then WLC 0131 PUS -79- the combined extracts are dried (Na2S04) and filtered to give crude 4-chloro-7-fluoropyrido [4, 3- d] yrimidine . 3-Bromoaniline (1.26 g, 7.35 mmole) , 3- bromoaniline hydrochloride (20 mg) and dry isopropanol (5 mL) are added, then the resulting solution is concentrated under vacuum to remove the CH2C12 and stirred at 20 °C for 1 h. Upon addition of dilute NaHC03 and water, the product crystallises. Filtration, washing with water and CH2C12, gives pure 4- (3- bromoanilino) -7-fluoropyrido [4, 3-d] pyrimidine (297 mg, • 67 %) as a cream solid. lK NMR (DMSO) δ 10.38 (1H, brs), 9.59 (1H, s) , 8.72 (1H, s) , 8.17 (1H, s) , 7.85 (1H, m) , 7.38 (3H, m) .

Example 13 7-Amino-4-anilinopyrido Γ4.3-dl pyrimidine 4.6-Diamino-2-bromo-3-cvanopyridine . HBr is bubbled for 2 h into a mixture of malononitrile (16.3 g, 0.247 mol) and toluene (400 mL) at O'C. A light yellow precipitate" forms. The reaction mixture is then heated at 100 *C for 2 h, with much gas evolution. After coolings to room temperature, the yellow solid is isolated via suction filtration, washed with toluene and air dried. The solid (25.96 g) is mixed with water (500 mL) , and the pH of the suspension is adjusted to 9 ~ 10 with NH40H (cone. ~ 15 mL) . After stirring at room temperature for 1 h, the mixture is filtered. Recrystallization from EtOH affords a yellow solid. After drying at 60 *C in a vacuum oven, 4 , 6-diamino-2-bromo-3-cyanopyridine (12.95 g, 49%) is obtained. 1H NMR (DMSO) δ 6.67 (2H,brs) , 6.55 (2H,brs) , 5.59 (lH,s) .

WLC 0131 PUS -80- 2 , -Diamino-5-cvanopyridinium acetate . 4,6-Diamino-2-bromo-3-cyanopyridine (12.77 g, 60 mmol) is hydrogenated in THF/MeOH (240 mL, 2:1) containing KOAc (5.9 g, 60 mmol) and 20% Pd/C (0.5 g) at 18 psi at 25 *C for 4 h. The mixture is celite filtered and the solvent is stripped under reduced pressure to give a solid (11.15 g) which is stirred with THF (100 mL) at room temperature for 20 min. The mixture is refiltered and the filtrate is stripped to dryness to give the desired product. After drying in a vacuum oven, 2 , 4-diamino-5-cyanopyridinium acetate (10.65 g, 92%) is collected as a yellow solid. H MR (DMSO) δ 7.90 (1H, s) , 6.26 (4H, brs) , 5.62 (1H, s) , 1.90 (3H, s). 7-Amino-4-thiono-3H-pyrido Γ4 , 3-dl pyrimidine .

A mixture of 2 , 4-diamino-5-cyanopyridinium acetate (0.199 g, 1.0 mmol), triethyl orthoformate (1.95 mL) and Ac20 (1.95 mL) is refluxed under N2 with stirring for 3 h. The solvent is then stripped and the residue is dissolved in ΜόΟΗ (10 mL) containing NaOMe (0.81 g, 15 mmol) . H2S is bubbled through the mixture for -5 min, which is then refluxed overnight. After the solvent is stripped, the residue is dissolved in hot water and boiled with charcoal. After filtration, the filtrate is neutralized with acetic acid whilst hot to generate a yellow solid. On cooling, the solid is collected by suction filtration, and is dried in a vacuum oven overnight. 7-Amino-4-thiono-3H- pyrido [4 , 3-d] pyrimidine (84 mg, 51%) is isolated as light yellow solid. *H NMR (DMSO) δ 9.82 (1H, s) , 9.34 (1H, s) , 8.37 (1H, s) , 7.80 (2H, d, J = 7.5 Hz), 7.38 (2H, t, J = 7.5 Hz), 7.12 (1H, t, J - 7.5 Hz) , 6.61 (2H, brs) 6.43 (1H, s) .

WLC 0131 PUS -81- 7-Amino-4-methylthiopyrido Γ4 , 3-dl pyrimidine . NEt3 (6 mL, 43 mmol) is added to a solution of 7-amino- 4-thiono-3H-pyrido [4, 3-d] yrimidine (0.77 g, 4.3 mmol) in DMSO (7 mL) stirred under N2 at 25°C. After the two phases have been stirred for 20 min, Mel (0.26 mL, 4.2 mmol) is added. After 2 h, the reaction mixture is poured onto stirring ice-water. Solid forms instantly. After further cooling at 0°C, the solid is collected by suction filtration and dried in a vacuum oven to give 7-amino-4-methylthiopyrido [4, 3-d] pyrimidine (0.564 g, 68%) . XH MR (DMSO) 6 8.98 (1H, s) , 8.71 (1H, s) , 6.94 (2H, brs) , 6.49 (1H, s) 2.63 (3H, s) . 7-Amino-4-anilinopyrido Γ4.3 -dl pyrimidine . A mixture of 7-amino-4-methylthiopyrido [ , 3-d] pyrimidine (0.136 g, 0.7 mmol) and aniline (0.5 mL, 5.5 mmol) is refluxed under N2 at 180°C for 2 h. The reaction mixture is cooled to 25°C, when it precipitates. The solid is collected by suction filtration and recrystallized from isopropanol, and dried in a vacuum oven overnight. 7-Amino-4-anilinopyrido [4 , 3- d] yrimidine (84 mg, 51%) is isolated as a light yellow solid.* *H MR (DMSO) δ 9.82 (1H, s) , 9.34 (1H, — s) , 8.37 (1H, s) , 7.80 (2H, d, J = 7.5 Hz), 7.38 (2H, t, J = 7.5 Hz), 7.12 (1H, t, J = 7.5 Hz) , 6.61 (2H, ' brs) 6.43 (1H, s) .

Example 14 7-Amino-4- (3-hydroxyanilino) pyrido f4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 - d] pyrimidine (299 mg, 1.56 mmole) and 3-aminophenol (1.60 g, 14.7 mmole) is stirred at 160 °C for 15 min. The resulting product is chromatographed over silica LC 0131 PUS -82- gel (9% MeOH/CH2Cl2) to give 7-amino-4- (3-hydroxyanilino) pyrido [4 , 3-d] yrimidine (108 mg, 18%) as a pale orange solid. 1H M (DMSO) δ 9.69 (1H, brs), 9.44 (1H, brs) , 9.33 (1H, s) , 8.38 (1H, s) , 7.37 (1H, t, J = 2.1 Hz), 7.21 (1H, brd, J = 8.4 Hz), 7.14 (1H, t, J = 8.0 Hz), 6.59 (2H, brs), 6.53 (1H, ddd, J = 7.9, 2.2, 0.8 Hz), 6.43 (1H, s) .

Example 15 7 -Amino-4 - ( 3 -methoxyanilino) yrido Γ4 , 3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3-d] pyrimidine (226 mg, 1.18 mmol) (described in the previous experimental) and m-anisidine (1.00 mL, 8.90 mmol) is stirred under N2 at 190 °C for 1.5 h. The resulting product is chromatographed over silica gel (5-7% EtOH/EtOAc) to give 7-amino-4- (3-methoxyanilino) pyrido [4 , 3-d] yrimidine (136 mg, 43%) as a light brown solid. ¾ NMR (DMSO) 5 9.IS (1H, brs), 9.34 (1H, s) , 8.40 (1H, s) , 7.50 (1H, brs), 7.44 (1H, d, J » 8.0 HZ) , 7.28 (1H, t, J - 8.2 Hz), 6.71 (1H, dd, J - 8.2, 2.3 Hz), 6.61 (2H, brs), 6.45 (1H, s) , 3.77 (3H,- s) .

Example 16 7-Amino-4 - (2-methoxyanilino) pyrido f4.3-dl yrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -d] pyrimidine (227 mg, 1.18 mmole) and o-anisidine (1.00 mL, 8.87 mmol) is stirred under N2 at 180 °C for 2.5 h. The resulting product is chromatographed over silica gel (5% EtOH/EtOAc) to give 7-amino-4- (2-methoxyanilino) pyrido [4 , 3-d] yrimidine (147 mg, 47%) as a yellow solid. XH NMR (DMSO) δ 9.44 (1H, brs), 9.25 WLC 0131 PUS -83- (1H, s) , 8.22 (1H, s), 7.54 (1H, dd, J = 7.7, 1.4 Hz), 7.24 (1H, ddd, J = 8.1, 7.4, 1.5 Hz) , 7.10 (1H, dd, J = 8.2, 1.2 Hz), 6.98 (1H, dt, Jd = 1.3 Hz, Jt = 7.5 Hz), 6.52 (2H, brs) , 6.41 (1H, s) , 3.79 (3H, s) .

Example 17 7-Amino-4 - (3-aminoanilino) pyrido f4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3-d] yrimidine (307 mg, 1.60 mmol) (described in a previous experimental) and 3-nitroaniline (2.00 g, 14.5 mmol) is stirred at 200 °C for 1.5 h, and the crude product is suspended in MeOH/THF (4:1, 250 rtiL) and hydrogenated over 5% Pd/C (2 g) at 60 psi and 20 °C for 24 h. The solution is filtered over celite, washing thoroughly (hot MeOH) , and is then absorbed onto alumina and chromatographed on alumina (4-8% EtOH/CHClj) to give 7-amino-4- (3-aminoanilino) pyrido [4 , 3-d] pyrimidine (66 mg, 16%) as a green solid,. Ή NMR (DMSO) δ 9.57 (1H, brs), 9.30 (1H, s) , 8.33 (1H, s),-¾7.04 (1H, t, J = 2.0 Hz), 6.99 (1H, t, J = 8.0 Hz), 6.88 (1H, brd, J = 8.0 Hz) , 6.55 (2H, brs), 6.40 (1H, s) , 6.34 (1H, dd, J = 7.9, 1.3 Hz), 5.10 (2H, brs) .

Example 18 7-Amino-4- (4-aminoanilino) yrido f4.3-dl pyrimidine 7-Amino-4 - (4 -acetamidoanilino) pyrido [4,3- dl yrimidine . A mixture of 7-amino-4- methylthiopyrido [4, 3-d] pyrimidine (138 mg, 0.72 mmolei · and 4-aminoacetanilide (1.50 g, 10.0 mmole) is stirred under N2 at 200 °C for 1 h. The resulting product is chromatographed over alumina (8-10% MeOH/CH2Cl2) to LC 0131 PUS -84- give 7-amino-4- (4-acetamidoanilino)pyrido [4 , 3-d] yrimidine (110 mg, 52%) as a pale yellow solid. 1H MR (DMSO) δ 9.94, 9.79 (1H, 1H, 2 brs) , 9.31 (1H, s) , 8.34 (1H, s) , 7.69 (2H, d, J = 8.9 Hz), 7.57 (2H, d, J = 8.9 Hz), 6.57 (2H, brs) , 6.43 (1H, s) , 2.05 (3H, s) . 7-Amino-4- (4 -aminoanilino) yrido Γ4 , 3-dl pyrimidine . A solution of 7-amino-4- (4-acetamidoanilino) pyrido [4, 3-d] pyrimidine (0.30 g, 1.02 mmole) in aqueous NaOH (2 M, 10 mL) and MeOH (10 mL) is stirred at 100 °C for 7 h. The resulting product is chromatographed over alumina (3-4% EtOH/CHCl3) to give 7-amino-4- (4-aminoanilino) pyrido [4 , 3-d] yrimidine (86 mg, 33%) as an orange solid. ¾ NMR (DMSO-) δ 9.58 (ΙΗ,' brs), 9.24 (1H, s) , 8.25 (1H, s) , 7.31 (2H d, J = 8.6 Hz), 6.58 (2H, d, J = 8.6" Hz), 6.48 (2H, brs), 6.39 (1H, s) , 5.00 (2H, brs) .

Example 19 7-Amino-4 - ( 3 -dimethviaminoanilino) pyrido f .3 -dl pyrimidine 4 A mixture of 7-amino-4-methylthiopyrido [4 , 3-d] pyrimidine 245 mg, 1.28 mmol) (described in a previous experimental) and N, IV-dimethy1-1, 3-phenylenediamine (1.60 g, 11.8 mmol) is stirred under N2 at 190 °C for 1 h, and the resulting product is chromatographed (twice) over alumina (3% EtOH/CHCl3) to give 7-amino-4- (3-dimethylaminoanilino)pyrido[4,3-d] pyrimidine (113 mg, 32%) as a pale yellow solid. XH NMR (DMSO) δ 9.66 (1H, brs) , 9.33 (1H, s) , 8.36 (1H, s), 7.22 (1H, brd, J = 7.8 Hz), 7.16 (2H, m) , 6.57 (2H, brs), 6.51 (1H, ddd, J = 8.0, 2.3, 1.2 Hz), 6.42 (1H, s) , 2.91 (6H, s) .

LC 0131 PUS -85- Example 20 7-Amino-4- (4-dimethylaminoanilino) pyrido Γ4 , 3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -d] pvrimidine (256 mg, 1.33 mmole) and iV, W-dimethyl-1, 4-phenylenediamine (1.95 g, 14.4 mmole) is stirred under N2 at 190 °C for 20 min. The resulting product is chromatographed over alumina (3-7% EtOH/CHCl3) to give 7-amino-4- (4-dimethylaminoanilino)pyrido [4,3-d]pyrimidine (198 mg, 53%) as an orange solid. *H NMR (DMSO) δ 9.67 (1H, brs) , 9.27 (1H, s) , 8.27 (1H, s) , 7.51 (2H, d, J = 8.9 Hz), 6.75 (2H, d, J = 8.9 Hz), 6.51 (2H, brs), 6.39 (1H, s) , 2.89 (6H, s) .

Example 21 7-Amino-4- (2-nitroanilino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3-dlpyrimidine (220 mg, 1.15 mmole) and 2-nitroaniline (2.00 g, 14.5 mmole) is heated to 100 °C, then excess dry HCl gas is added to the hot stirred solution, and the mixture stirred at 160 °C for 20 min. The resulting product is neutralized with excess NaHC03, dissolved in MeOH/CHCl3, dried onto silica gel and chromatographed over silica gel (2-4% MeOH/CH2Cl2) to give 7-amino-4- (2-nitroanilino) pyrido [4, 3-d] pyrimidine (108 mg, 33%) as a yellow brown solid. Ή NMR (DMSO) δ 10.40 (1H, brs), 9.24 (1H, brs), 8.20 (1H, brs), 8.12 (1H, brs), 8.01 (2H, brs), 7.75 (1H, brs), 6.70 (2H, brs) , 6.43 (1H, brs) .

WLC 0131 PUS -86- Example 22 7-Amino-4- (3 -nitroanilino) pyrido [ , 3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3-d]pyrimidine (127 mg, 0.66 mmol) (described in a previous experimental) and 3-nitroaniline (1.70 g, 12.3 mmol) is stirred under N2 at 200°C for 1.5 h. The resulting product is chromatographed over alumina (5-20% EtOH/CHCl3) to give 7-amino-4- (3 -nitroanilino) pyrido [4, 3-d] pyrimidine (81 mg, 39%) as a brown solid. XH NMR (DMSO) δ 10.17 (1H, brs) , 9.37 (1H, s) , 8.87 (1H, brs), 8.48 (1H, s) , 8.33 (1H, brd, J = 7.5 Hz), 7.95 (1H, ddd, J = 8.2, 2.1, 1.0 Hz), 7.67 (1H, t, J = 8.2 Hz), 6.70 (2H, brs) , 6.47 (1H, s) .

Example 23 7-Amino-4- ( 3 - fluoroanilino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -d] pyrimidine (215 mg, 1.12 mmol) and 3-fluoroaniline (1.16 g, 10.4 mmo?) is stirred at 160 °C for 30 min. The resulting product is chromatographed over silica gel (6-7% MeOH/CHzCl2) to give 7-amino-4- (3- fluoroanilino) pyrido [4 , 3 -d] pyrimidine (185 mg, 65%) as a white solid. XH NMR (DMSOJ δ 9.94 (1H, brs), 9.36 (1H, s) , 8.46 (1H, s) , 7.91 (1H, brd, J = 11.9 Hz), 7.63 (1H, brd, J = 8.1 Hz), 7.41 (1H, dd, J « 15.7, 7.7 Hz), 6.93 (1H, dt, Jt = 8.5 Hz, Jd = 2.4 Hz), 6.68 (2H, brs) , 6.38 (1H, s) .

LC 0131 PUS -87- Example 24 7-Amino-4- ( 3 -chloroanilino) pyrido [4 , 3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -d]pyrimidine (208 mg, 1.08 mmol) and 3-chloroaniline (1.21 g, 9.48 mmol) is stirred at 150 °C for 20 min. The resulting product is chromatographed over alumina (5-10% MeOH/CH2Cl2) to give 7-amino-4- (3-chloroanilino) pyrido [4 , 3-d] pyrimidine (177 mg, 60%) as a white solid. Ή N R (DMSO) δ 9.92 (lH,brs), 9.35 (1H, s) , 8.45 (1H, s) , 8.08 (1H, brs), 7.79 (1H, brd, J = 8.0 Hz), 7.40 (1H, t, J = 8.1 Hz), 7.16 (1H, dd, J = 7.9, 1.3 Hz), 6.68 (2H, brs), 6.46 (1H, s) .

Example 25 7-Amino-4 - (3.4 -dichloroanilino) yrido Γ4.3-dl pyrimidine A mixture of 7-amino-4 -methylthiopyrido [4 , 3 -d] yrimidine (247 mg, 1.29 mmol) and 3,4-dichloroaniline (1.50 g, 9.26 mmol) is stirred at 165 °C for 30 min. The^ resulting product is chromatographed over silica gel (7-8% MeOH/CH2Cl2) to give 7-amind—1- (3 , -dichloroanilino) pyrido [4 , 3-d] pyrimidine (252 mg, 64%) as a pale yellow solid. XH NMR (DMSO) δ 9.97 (1H, brs),.9.34 (1H, s) , 8.47 (1H, s) , 8.29 (1H, brs), 7.86 (1H, brd, J = 8.6 Hz), 7.62 (1H, d, J = 8.8 Hz), 6.70 (2H, brs), 6.46 (1H, s) .

Example 26 7-Amino-4- ( 2 -bromoanilino) pyrido f4.3-dl pyrimidine A mixture of 7-amino-4 -methylthiopyrido [4 , 3 -d] yrimidine (198 mg, 1.03 mmol) (described in a previous experimental) and 2-bromoaniline (1.00 mL, WLC 0131 PUS -88- 9.18 mmol) is stirred under N2 at 180°C for 2.5 h, and the resulting product is chromatographed on alumina (1% EtOH/CHCl3) to give 7-amino-4- (2-bromoanilino) pyrido [4, 3-d] pyrimidine (108 mg, 33%) as a pale yellow solid, *H MR (DMSO) δ 9.91 (1H, brs) , 9.27 (1H, s) , 8.20 (1H, s) , 7.73 (1H, d, J = 7.9 Hz), 7.50 (1H, m) , 7.44 (1H, t, J = 6.9 Hz), 7.25 (1H, m) , 6.59 (2H, brs), 6.42 (1H, s) .

Example 27 7-Amino-4- (3 -bromoanilino) pyrido 14.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4, 3-d] pyrimidine (167 mg, 0.87 mmol) (described in a previous experimental) and 3-bromoaniline (0.75 mL, 7.8 mmol) is stirred under N2 at 190°C for 2.5 h, and the precipitate which appears on cooling is recrystallized from Pr^H. XH NMR (DMSO) δ 9.91 (1H, brs), 9.34 (1H, s) , 8.45 (1H, s) , 8.19 (1H, s) , 7.84 (1H, d, J = 8.0 Hz) / 7.34 (1H, t, J = 8.0 Hz), 7.29 (1H, d, J = 8.2 Hii) , 6.68 (2H, brs) , 6.45 (1H, s) .

Example 28 7-Amino-4 - (4 -bromoanilino) yrido f4 ■ 3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 - d] pyrimidine (261 mg, 1.36 mmole) and 4-bromoaniline (1.00 g, 5.81 mmole) is stirred under N2 at 200 °C for 15 min. The resulting product is chromatographed on silica gel (10-15% EtOH/EtOAc) to give 7-amino-4- (4- bromoanilino) yrido [4 , 3 -d] pyrimidine (200 mg, 46%) as a pale yellow solid. *H NMR (DMSO) δ 9.88 (1H, brs), 9.34 (1H, s) , 8.40 (1H, s) , 7.83 (2H, d, J = 8.8 Hz), LC 0131 PUS -89- 7.55 (2H, d, J = 8.8 Hz), 6.64 (2H, brs) , 6.44 (1H, s) .

Example 29 7-Amino-4- (3-iodoanilino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [ , 3 -d] pyrimidine (72 mg, 0.37 mmol) and 3-iodoaniline (1.25 g, 5.71 mmol) is stirred at 160 °C for 30 min. The resulting product is chromatographed over silica gel (5-7% MeOH/CH2Cl2) to give 7-amino-4- (3 -iodoanilino) pyrido [ , 3 -d3 pyrimidine (83 mg, 61%) as a light brown rosettes. :H NMR (DMSO) δ 9.84 (1H, brs), 9.34 (1H, s) , 8.44 (1H, s) , 8.30 (1H, brs) , 7.90 (1H, ' dd, J = 7.9, 0.8 Hz), 7.47 (1H, d, J - 7.7 Hz) , 7.18 (1H, t, J = 8.0 Hz), 6.66 (2H, brs), 6.46 (1H, s) .

Example 30 7-Amino-4- (2- rifluoromethylanilino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -d] pyrimidine (300 mg, 1.56 mmol), 2-aminobenzotri-fluoride hydrochloride (1.00 g, 5.06 mmol) and 2-aminobenzotrifluoride (2.00 g, 12.4 mmol) is stirred at 160 °C for 10 min. The resulting product is neutralized with excess NaHC03, dissolved in MeOH/CHCl3, dried onto silica gel and chromatographed over silica gel (6-7% MeOH/CH2Cl2) to give 7-amino-4- (2-trifluoro-methylanilino) pyrido [4 , 3-d] pyrimidine (194 mg, 41%) as a cream solid, mp (MeOH/CHCl3/light petroleum) 126-130 °C (dec). XH NMR (DMSO) δ 10.60 (1H, brs), 9.17 (1H, brs), 8.13 (1H, brs), 7.76, 7.69 (1H, 1H, m, m) , 7.45 (2H, m) , 6.66 (2H, brs), 6.36 (1H, s) .

WLC 0131 PUS -90- Example 31 7 -Amino- - ( 3 - trifluoromethylanilino) pyrido [4.3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3-d]pyrimidine (234 mg, 1.22 mmol) (described in a previous experimental) and 3-aminobenzotrifluoride (2.00 mL, 16.0 mmol) is stirred under N2 at 190-200°C for 2 h, and the resulting product is then chromatographed over silica gel (5-10% EtOH/EtOAc) , and then over alumina (5-7% EtOH/CHCl3) to give 7-amino-4- (3 -trifluoromethylanilino) pyrido [4 , 3-djpyrimidine (157 mg, 42%) as a cream solid. ·¾ NMR (DMSO) b 10.04 (1H, s) , 9.37 (1H, s) , 8.46 (1H, s) , 8.31 (1H, s) , 8.19 (1H, d, J * 8.2 Hz), 7.62 (1H, t, J = 8.0 HZ), 7.45 (1H, d, J = 7.7 Hz), 6.69 (2H, brs) , 6.47 (1H, s) .

Example 32 7-Amino-4- (4- rifluoromethylanilino) pyrido Γ4.3-dl pyrimidine A mixtuie of 7 -amino-4 -methylthiopyrido [4 , 3-djpyrimidine (390 mg, 2.03 mmol), 4-aminobenzotri-fluoride hydrochloride (0.40 g, 2.02 mmol) and 4-aminobenzotrifluoride (1.61 g, 10.0 mmol) is stirred at 180 °C for 2 min. The resulting product is neutralized with excess NaHC03, dissolved in MeOH/CHCl3< dried onto alumina and chromatographed over alumina (4-7% MeOH/CH2Cl2) to give 7-amino-4- (4-trifluoromethylanilino) pyrido [4 , 3 -d] pyrimidine (390 mg, 63%) as a cream solid. Analytically pure material was obtained by further chromatography over silica gel (5% MeOH/CH2Cl2) to give pale yellow needles. XH NMR (DMSO) b 10.09 (1H, brs), 9.40 (1H, s) , 8.48 (1H, s) , 8.13 (2H, d, J = 8.2 Hz), 7.74 (2H, d, J = 8.7 Hz), 6.72 (2H, brs) , 6.40 (1H, s) .

WLC 0131 PUS -91- Example 33 4- (3 -Bromoanilino) -7-methylaminopyrido Γ4 , 3-d] pyrimidine A mixture of 7-fluoro-4- (3 -bromoanilino) -pyrido [4, 3-d] yrimidine (74 mg, 0.23 mmol) , triethyl-amine (7 mL, 50 mmol) and methylamine hydrochloride (3.0 g, 44 mmol) in isopropanol (30 mL) contained in a steel bomb is stirred at 95 °C (oil bath) for 5 h.

The resulting mixture is concentrated under vacuum, basified with aqueous Na2C03, diluted with water and extracted with EtOAc (3 x 100 mL) . Chromatography of this extract on silica gel (3 % MeOH/CH2Cl2) gives 4- ( 3 -bromoanilino) - 7-methylaminopyrido [4 , 3 -d] pyrimidine (50 mg, 65%) as a pale yellow solid. lH NMR (DMSO) δ ' 9.93 (1H, brs) , 9.37 (1H, s) , 8.47 (1H, s) , 8.18 (1H, s) , 7.84 (1H, d, J = 7.8 Hz) , 7.34 (1H, t, J = 7.9 Hz), 7.30 (1H, brd, J = 8.1 Hz) , 7.19 (1H, q, J = 4.7 Hz), 6.35 (1H, s) , 2.85 (3H, d, J = 4.8 Hz).

Example 34 4- (3-Bromoanilino) -'■7-dimethylaminopyrido Γ4.3 -dl pyrimidine A njixture of 7-fluoro-4- (3 -bromoanilino) pyrido [4 , 3-d] pyrimidine (101 mg, 0.32 mmol), triethylamine (4.4 mL, 32 mmole) and dimethylamine hydrochloride (2.58 g, 32 mmol) in isopropanol (30 mL) contained in a steel bomb is stirred at 100 °C (oil bath) for 4 h. The resulting solution is concentrated under vacuum, basified with aqueous Na2C03 and diluted with water to give a solid. Filtration and recrystallisation from MeOH/CHCl3 gives 7-dime hylamino-4 - ( 3 -bromoani1ino) yrido [4,3- d]pyrimidine (102 mg, 94%) as a pale yellow solid. -H NMR (DMSO) δ 9.93 (1H, brs) , 9.42 (1H, s) , 8.48 (1H. s) , 8.19 (1H, s) , 7.85 (1H, d, J = 7.7 Hz) , 7.35 CH, WLC 0131 PUS -92- t, J = 7.9 Hz), 7.30 (1H, brd, J = 7.8 Hz), 6.53 (1H, s) , 3.16 (6H, s) .

Example 35 4- ΓΝ- (3-Bromophenyl) -N-methylaminol -7-methylamino-pyrido f4 , 3 -dl pyrimidine A mixture of 7-fluoro- - (3-bromoanilino) -pyrido [4 , 3-d] pyrimidine (100 mg, 0.31 mmole) , trieth-ylamine (4.4 mL, 32 mmole) and methylamine hydrochloride (2.12 g, 32 mmole) in isopropanol (30 mL) con-tained in a steel bomb is stirred at 100 °C (oil bath) for 5 h. The resulting mixture is concentrated under vacuum, basified with aqueous Na2C03, diluted with water and extracted with EtOAc (3 x 100 mL) . Chromatography of this extract on silica gel (1-2% eOH/CH2Cl2) gives 4- [N- (3-bromophenyl) -N-methylamino] -7-methylaminopyrido [4, 3-d] pyrimidine (23 mg, 21%) as a pale yellow solid. XH N R (DMSO) δ 8.14 (1H, s) , 7.79 (1H, s) , 7.30 (1H, t, J = 8.0 Hz), 7.20 (1H, ddd, J = 7.9, 1.8, 0.8 Hz) ,"' 7.03 (1H, brq, J" = 4.9 Hz) , 7.01 (1H, t, J ■ 1.9 Hz), 6.82 (1H, ddd, J = 7.8, 1.8, 0.9 Hz) , 6.25 (1», s) , 3.40 (3H, s) , 2.73 (3H, d, J = 4.9 Hz) .

Example 36 7-Acetylamino-4- (3 -bromoanilino) pyrido Γ4.3 -dl pyrimidine A mixture of 7-amino-4- (3-bromoanilino) -pyrido [4, 3-d] pyrimidine (0.154 g, 0.49 mmol) , acetic anhydride (0.14 mL, 1.5 mmol), triethylamine (0.14 mL, 1.0 mmol) and a catalytic amount of 4- (NjN-dimethyl-amino) pyridine are stirred under N2 at room temperature for 18 h. The reaction is then quenched by addition of ice water. The dark precipitate is collected by WLC 0131 PUS -93- Buchner filtration and is purified by preparative tic (Rf = 0.25, 7% MeOH/CHCl3) . Recrystallization from EtOH gives 7-acetylamino-4- (3 -bromoanilino) pyrido [4 , 3 -d] -pyrimidine (13.5 mg, 7.7%). XH NMR (DMSO) δ 10.92 (1H, S) , 10.22 (1H, s) , 9.64 (1H, s) , 8.70 (1H, s) , 8.28 (1H, s), 8.21 (lH,s), 7.88 (1H, d, J = 7.7Hz) 7.41-7.34 (3H, m) , 2.16 (3H, s) .

Example 37 4- (3 -Bromoanilino) -7-methoxypyrido Γ4.3 -dl pyrimidine A solution of 7-fluoro-4- (3 -bromoanilino) -pyrido [4, 3-d] pyrimidine (100 mg, 0.31 mmol) in 1 M sodium methoxide-methanol (30 mL) is stirred under reflux for 42 h. The resulting mixture is concentrated under reduced pressure, diluted with water and neutra-lized with dilute HCl to give 7-methoxy-4- (3 -bromoanilino) pyrido [4 , 3 -d] pyrimidine (92 mg, 89%) as a white solid. XH NMR (DMSO) δ 10.22 (1H, brs) , 9.57 (1H, S) , 8.63 (1H, s) , 8.19 (1H, s) , 7.86 (1H, brd, J = 7.9 Kz), 7.39 (1H, t, ' J = 7.9 Hz), 7.35 (1H, dd, J = 7.9, 1.5 Hz), 6.96 (1H, s) , 4.00 (3H, s) .

Example 38 4-Benzylaminopyrido Γ4.3 -dl yrimidine 4 -Methylthiopyrido [4, 3-d] pyrimidine (160.4 mg, 0.902 mmol), and benzylamine (106.3 mg, 0.992 mmol) in EtOH (2 mL) are heated at 80°C for 12 h, and then the solvent is removed under reduced pressure . The resulting solid is suspended in CH2C12, filtered, and the resulting solid is purified by preparative tic on silica, eluting with 5% MeOH in CHC13. Removal of the solvent under reduced pressure yields 4- WLC 0131 PUS -94- benzylaminopyrido [4, 3-d] pyrimidine (36 mg, 17%) . ικ NMR (DMSO) δ 9.60 (1H, s) , 9.37 (1H, t, J = 5.8 Hz), 8.72 (1H, d, J = 5.8 Hz), 8.57 (1H, s) , 7.54 (1H, d, J = 5.8 Hz), 7.37 (2H, d, J = 7.0 Hz), 7.33 (2H, t , J = 7.3 Hz), 7.25 (1H, t, J=7.2 Hz), 4.81 (2H, d, J = 5.8 Hz) .

Example 39 4- ( fRl -l-Phenylethylamino) pyrido Γ4.3-d] pyrimidine To a mixture of 4-methylthiopyrido [4 , 3-d] pyrimidine (85 mg, 0.48 mmol) and EtOH (2.5 mL) is added R-methylbenzylamine (0.13 mL, 1.0 mmol) dropwise. The resulting mixture is refluxed at 80°C for 20 h. The solvent is removed under reduced pressure to give an oil which is crystallized from MeOH to give 4 - ( [R] -1-phenylethylamino) yrido [4 , 3 -d] pyrimidine (41.6 mg, 35%), mp 138-138.5°C. ¾ NMR (DMSO) δ 9.77 (1H, d, J = 0.7 Hz), 9.00 (1H, d, J = 7.7 Hz), 8.73 (1H, d, J = 5.8 Hz), 8.54 (1H, s) , 7.53 (1H, dd, J = 5.8, 0.5 Hz), 7.45 (2H, d, J = 7.2 Hz), 7.33 (2H, t, J = 7.6 Hz) , 7.23 (1H, tt, J = 7.5, 1.2 Hz), 5.63 (1H., p, J = 7.2 Hz), 1.61 (3H, t, J = 7.0 Hz) .

Example 40 7-Amino-4 -benzylaminopyrido Γ4 , 3 -dl pyrimidine A mixture of 2 , 4 -diamino, 5-cyanopyridinium acetate (8.78 g, 45 mmol), formic acid (10.66 g, 0.204 mol) and benzylamine (45 mL, 0.41 mol) is heated at 200 *C under N2 for 2 h. Upon cooling, it solidifies. Water (500 mL) is added and the gummy solid/water mixture is stirred for -20 min. at 0*C. The liquid is WLC 0131 PUS -95- decanted. The solid is washed with water and then recrystallized from isopropanol (25 mL) . After drying in a vacuum oven overnight, 7-amino-2-benzylaminopyrido [4, 3-d] pyrimidine (8.29 g, 73%) is obtained as a light yellow solid. XH NMR (DMSO) δ 9.10 (1H, S) , 8.85 (1H, t, J = 5.8 Hz) , 8.25 (1H, s) , 7.21-7.36 (5H, m) , 6.46 (2H, brs) , 6.35 (1H, s) , 4.74 (2H, d, J = 6.0 Hz) .

Example 41 7-Amino-4- ( iRl - 1-phenylethylamino) pyrido [4 , 3 -dl pyrimidine A mixture of [R] -1-phenylethylamine (0.072 mL, 0.55 mmol) and 7-amino-4-methylthiopyrido [4 , 3 -d] pyrimidine ( 97 mg, 0.5 mmol) (described in a previous experimental) is heated at 180°C under N2 for 1.5 hr. The reaction is then cooled to room temperature producing a precipitate. The mixture is added to water and CHC13> sonicated and filtered. The phases are separated and the aqueous phase is extracted with CH 13. The combined extracts are washed with water, saturated brine and dried (MgS04) . The solvent is removed under reduced pressure and the residue purified by using preparative tic (5% MeOH/CHClj) and recrystallization from CHC13 to give 7-amino-4- ( [R] -1-phenylethylamino) pyrido [4 , 3-d] pyrimidine (14.5 mg, 11%) , mp 231.8-232.1°C. XH NMR (DMSO) δ 9.23 (1H, s) , 8.50 (1H, d, J = 8.0 Hz), 8.19 (1H, s) , 7.41 (2H( d, J = 7.0 Hz), 7.31 (2H, t, J = 8.0 Hz, 7.21 (1H, tt, J = 7.4, 1.2 Hz), 6.45 (2H, s) , 6.33 (1H, s) , 5.56 (1H, p, J = 7.2 Hz), 1.55 (3H, d, J = 7.0 Hz) .

WLC 0131 PUS -96- Example 42 7-Amino-4- (2 -aminobenzylamino) pyrido Γ4.3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 d] pyrimidine (136 mg, 0.71 mmol) (described in a previous experimental) and 2-aminobenzylamine (1.70 g 13.8 mmol) in isopropanol (5 mL) is stirred at reflux for 1 h, and the resulting product is chromatographed on silica gel (7-20% EtOH/EtOAc) and alumina (6-10% EtOH/CHCl3) to give 7-amino-4- (2-aminobenzylamino) pyrido [4 , 3-d] pyrimidine (89 mg, 47%) as a white solid. XH MR (DMSO) δ 9.08 (1H, s) , 8.68 (1H, t, J - 5.8 Hz), 8.26 (1H, s) , 7.05 (1H, d, J = 7.4 Hz), 6.96 (1H, t, J = 7.6 Hz), 6.63 (1H, d, J = 7.9 Hz), 6.51 (1H, t, J = 7.4 Hz), 6.46 (2H, brs) , 6.35 (1H, s) , 5.20 (2 H, brs), 4.56 (2H, d, J = 5.8 Hz) .

Example 43 7-Amino-4- ( 3 -dimethylaminobenzylamino) pyrido f4, 3 -dl -pyrimidine A isixture of 7-amino-4-methylthiopyrido [4 , 3 d] pyrimidine (236 mg, 1.23 mmol) (described in a previous experimental) and 3 -dimethylamino-benzylamine (1.36 g, 9.07 mmol) in isopropanol (5 mL) is stirred under N2 at reflux for 1 h, and the resulting product is chromatographed on silica gel (10-15% EtOH/EtOAc) , then on alumina (1% EtOH/CHCl3) to give 7-amino-4 - ( 3 - dimethylaminobenzylamino) pyrido [4 , 3 -d] pyrimidine (145 mg, 40%) as a white solid. XH NMR (DMSO) δ 9.11 (1H, S) , 8.79 (1H, t , J = 5.9 Hz), 8.26 (1H, s) , 7.11 (1H. dd, J = 8.0, 7.7 Hz), 6.73 (1H, brs), 6.63 (1H, d, J 7.6 Hz), 6.60 (1H, dd, J = 8.1, 2.2 Hz), 6.44 (2H, LC 0131 PUS -97- brs) , 6.35 (1H, s) , 4.67 (2H, d, J = 5.8 Hz), 2.86 (6H, s) .

Example 44 7-Amino-4 - (3 -nitrobenzylamino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -djpyrimidine (228 mg, 1.19 mmol) (described in a previous experimental) and 3-nitrobenzylamine (0.81 g, 5.33 mmol) is stirred under N2 at 150-160 °C for 1.5 h, and the resulting product chromatographed on silica gel (5-10% EtOH/EtOAc) to give 7-amino-4- (3-nitrobenzylmino)pyrido[4,3-d]pyrimidine (151 mg, 43%) as a yellow solid. XH NMR (DMSO) δ 9.11 (1H, s) , 8.98 (1H, t, J = 5.5 Hz), 8.26 (1H, s) , 8.22 (1H, brs) , 8.12 (1H, dd, J = 8.0, 1.8 Hz), 7.83 (1H, d, J = 7.7 Hz), 7.63 (1H, t, J = 7.9 Hz), 6.50 (2H, brs) , 6.38 (1H, s) , 4.85 (2H, d, J = 5.8 Hz) .

Example 45 7-Amino-4- (3-methox benzylamino)pyrido Γ4.3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3-d]pyrimidine (136 mg, 0.71 mmol) (described in a previous experimental) and 3-methoxybenzylamine (1.37 g, 10.0 mmol) in isopropanol (3 mL) is stirred under N: at reflux for 3 h. Evaporation of the solvent and chromatography on silica gel (5-10% EtOH/EtOAc) gives 7-amino-4- (3-methoxybenzylamino)pyrido[4, 3-dlpyrimidine (153 mg, 77%) as a white solid. XH NMR (DMSO) δ 9.11 (1H, s) , 8.83 (1H, t, J = 5.7 Hz), 8.26 (1H, s) , 7.24 (1H, dt, Jd = 0.8 Hz, Jt = 8.1 Hz), 6.92 (2H, m) , 6.81 (1H, dt, Jd = 8.2 Hz, Jt =1.2 Hz), 6.46 (2H, brs), 6.37 (1H, s) , 4.71 (2H, d, J = 5.8 Hz), 3.73 (3H, s) .

LC 0131 PUS -98- Example 46 7-Amino-4- (4-chloroben2ylamino) pyrido Γ4 , 3 -dl pyrimidine mesylate The free base (56 mg, 0.20 mmol) (prepared from 2,4-diamino, 5-cyanopyridinium acetate, formic acid and 4-chlorobenzylamine at 200°C as described in a previous example is precipitated from acetone solution with methanesulfonic acid (105 /ili, 0.23 mmol) to give a polymesylate salt. XH NMR (DMSO) δ 10.59 (1H, t , J = 5.6 Hz), 9.24( 1H, s) , 8.69 (1H, s) , 7.42 (4H, s) , 6.42 (1H, s) , 5.8 (-6H, vbrs) , 4.89 (2H, d, J = 5.8 Hz) , 2.41 (-7.5H, s) .

Example 47 7-Amino-4- (2-bromobenzylamino) yrido [ .3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -dlpyrimidine (225 mg, 1.17 mmol) (described in a previous experimental) and 2-bromobenzylamine (0.84 g, 4.52 mmol) is stirred under N2 at 140 °C for 1 h, and the resulting product chromatographed on silica gel (1-5% EtOH/EtOAc) to give 7-amino-4- (2-bromobenzylamino) pyrido [4 , 3 -d] pyrimidine (175 mg, 45%) as a light brown solid. LH NMR (DMSO) b 9.16 (1H, s) , 8.85 (1H, t , J = 5.7 Hz), 8.24 (1H, s) , 7.64 (1H, d, J = 7.8 Hz), 7.34 (1H, dd, J = 7.7, 7.1 Hz), 7.31 (1H, dd, J = 7.7, 2.4 Hz), 7.21 (1H, ddd, J = 7.8, 6.9, 2.4 Hz), 6.50 (2H, brs) , 6.39 (1H, s) , 4.74 (2H, d, J = 5.7 Hz) .

WLC 0131 PUS -99- Example 48 7-Amino-4- (3 -bromobenzylamino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino- -methylthiopyrido [4 , 3 -d] pyrimidine (228 mg, 1.19 mmol) (described in a previous experimental) and 3-bromobenzylamine (0.84 g, 4.52 mmol) is stirred under N2 at 140°C for 1 h. The resulting product is chromatographed on silica gel (2-10% EtOH/EtOAc) to give 7-amino-4- [ (3-bromophenyl) methylamino] pyrido [4 , 3-d] pyrimidine (203 mg, 52%) as a light brown solid. XH R (DMSO) δ 9.09 (1H, S) , 8.86 (1H, t, J = 5.8 Hz) , 8.26 (1H, s) , 7.54 (1H, s) , 7.44 <1H, d, J = 7.8 Hz,), 7.36 (1H, d, J = 7.6 Hz), 7.29 (1H, t, J = 7.7 Hz) , 6.48 (2H, s) , 6.37* (1H, s) , 4.73 (2H, d, J = 5.8 Hz).

Example 49 7-Amino-4 - (4 -bromobenzylamino) yrido Γ4.3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 -d] pyrimidine (234 'mg, 1.22 mmol) (described in a previous experimental) and 4-bromobenzylamine (0.84 g, 4.52 mmol) is stirred under N2 at 140 °C for 1 h, and the resulting product chromatographed on silica gel (10% EtOH/EtOAc) to give 7-amino-4- (4-bromobenzylamino) pyrido [4, 3-d] pyrimidine (192 mg, 48%) as a cream solid. *H NMR (DMSO) δ 9.09 (1H, s) , 8.87 (1H, t, J = 5.7 Hz), 8.25 (1H, s) , 7.51 (2H, d, J = 8.3 Hz), 7.31 (2H, d, J = 8.3 Hz), 6.46 (2H, brs) , 6.37 (1H, s) , 4.70 (2H, d, J = 5.8 Hz).

WLC 0131 PUS -100- Example 50 7-Amino-4- (2 -trifluoromethylbenzylamino) pyrido Γ4 , 3 -dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 d] yrimidine (225 mg, 1.17 mmol) and 2- (trifluoromethyl) benzylamine (0.90 mL, 6.42 mmol) is stirred under N2 at 150 °C for 1 h. The resulting product is chromatographed on silica gel (5% EtOH/EtOAc) to give 7-amino-4- (2-trifluoromethylbenzyl) aminopyrido [4 , 3-d] yrimidine (0.22 g, 59V) as a white solid. XH MR (DMSO) δ 9.16 (1H, s) , 8.88 (1H, t, J = 5.7 Hz), 8.23 (1H, s) , 7.75 (1H, d, J = 7.7 Hz), 7.62 (1H, t, J = 7.5 Hz), 7.50 (1H, d, J = 7.4 Hz), 7.47 (1H, t, J = 7.6 Hz), 6.51 (2H, brs) , 6.39 (1H, s) , 4.92 (2H, d, J = 5.5 Hz).

Example 51 7 -Amino-4- (3 -trifluoromethylbenzylamino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 d] pyrimidine *(225 mg, 1.17 mmole) and 3- (trifluoromethyl) benzylamine (0.63 mL, 4.40 mmole) is stirred under N2 at 140 °C for 1 h. The resulting product is chromatographed on silica gel (3-5% EtOH/EtOAc) to give 7-amino-4- [ (3- trifluoromethylphenyl) methylamino] pyrido [4,3- d] pyrimidine (0.24 g, 63%) as a light brown solid. XH NMR (DMSO) δ 9.10 (1H, s) , 8.92 (1H, t, J = 5.7 Hz), 8.26 (1H, s) , 7.71 (1H, s) , 7.66 (1H, d, J = 7.4 Hz), 7.62 (1H, d, J = 7.8 Hz), 7.57 (1H, t, J = 7.6 Hz), 6.49 (2H, brs), 6.38 (1H, s) , 4.82 (2H, d, J = 5.8 Hz) . -101- Example 52 7-Amino-4- ( -trifluoromethylbenzylamino) pyrido [4, 3-d] pyrimi dine A mixture of 7-amin.o-4-methylthiopyrido [ 4, 3-d] pyrimidine (225 mg, 1.17 mmol) and 4- (trifluorornethyl ) benzylamine (0.63 L, 4.42 mmol) is stirred under N2 at 140°DC for 1 h. The resulting product is chromatographed on alumina (5-10% EtOH/CHCl3) then silica gel (2-10% EtOH/EtOAc) to give 7-amino-4- [ ( 4-trifluoromethylphenyl ) ethylamino] pyrido [4,3-d]pyrimidine (0.21 g, 56%) as a light brown solid. 1H NMR (DMSO) δ 9.12 (1H, s), 8.94 (1H, t, J = 5.8 Hz), 8.24 (1H, s), 7.69 (2H, d, J = 8.1 Hz), 7.56 (2H, d, J = 8.1 Hz), 6.48 (2H, brs), 6.38 (1H, s), 4.82 (2H, d, J = 5.8 Hz).

Example 53 7-Amino-4- ( ( thien-2-ylmethyl ) amino) pyrido [4, 3-d] pyrimidine dimesylate The compound is obtained from 2, 4-diamino, 5-cyanopyridinium acetate (190 mg, 0.98 mmol), formic acid (0.23 g, 4.4 mmol) and (thienylmethyl) amine (1.07 ml, 10 mmol) as described in a previous experimental. The crude product is converted into a dimesylate salt as described previously and recrystallized from Pr^OH to give 7-amino-4- (thien-2-ylmethylamino) pyrido [4, 3-d] pyrimidine dimesylate in 19% yield. 1H NMR (DMSO) δ 10.67 (1H, t, J = 5.8 Hz), 9.21 (1H, s) , 8.77 (1H, s), 7.48 (1H, dd, J = 5.1, 1.2 Hz) 7.16 (1H, dd, J = 3.4, 0.7 Hz), 7.02 (1H, dd, J = 4.8, 3.4 Hz), 6.42 (1H, s) , 5.06 (2H, d, J = 5.7 Hz), 2.41 (6H, s) .

LC 0131 PUS -102- Example 54 7 -Acetylamino-4 -benzylaminopyrido [4 , 3 -dl pyrimidine 7 -Acetylamino-4 -methylthiopyrido [4,3-dl pyrimidine . Acetyl chloride (0.70 mL, 9.84 mmol) is added to a solution of 7-amino-4 -methylthiopyrido [4 , 3 -d] pyrimidine (0.20 g, 1.04 mmol) (described in a previous experimental) and Et3N (1.51 mL, 10.8 mmol) in THF at 0°C, and then the mixture is stirred at 20 °C for 4 h. Water (50 mL) was added, then the solution was extracted with EtOAc (3 x 50 mL) . Evaporation and chromatography on alumina (1% EtOH/CHCl3) yields 7-acetylamino-4 -methylthiopyrido [4 , 3 -d] pyrimidine (0.12 g, 49%) as a yellow solid,. H MR (DMSO)- δ 11.05 ( ltf, s) , 9.30 (1H, s) , 9.02 (1H, s) , 8.38 (1H, s) , 2.71 (3H, s) , 2.18 (3H, s) . 7-Acetylamino-4 -benzylaminopyrido Γ4 , 3-dl pyrimidine . A mixture of 7-acetylamino-4 -methylthiopyrido [4 , 3-d] pyrimidine (0.40 g, 1.71 mmol) and benzylamine (5.0 mL, 9.15 mmol) is stirred under N2 at 140°C for 1 h, and the resulting product is chromatography on silica gel (EtOAc) to give 7-acetylamino-4-benzylaminopyrido [4, 3-d] pyrimidine (0.31 g, 62%) as a white solid. XH NMR (DMSO) δ 10.79 (1H, s) , 9.42 (1H, s) , 9.23 (1H, t, J = 5.8 Hz), 8.49 (1H, s) , 8.18 (1H, s) , 7.39 (1H, dt , Jd = 6.9 Hz, Jc =1.7 Hz), 7.34 (1H, tt, J = 7.3, 1.7 Hz) , 7.25 (1H, tt, J = 7.1, 1.7 Hz), 4.80 (2H, d, J = 5.8 Hz) , 2.15 (3H, s) . -103- Example 55 4-Anilinopyrido[3.4-d]pyrimidine 4-Carbamoylnicotinic acid. 3,4-Pyridine dicarboxylic anhydride (8.3 g, 55.6 mmol) is added to cone. NH4OH (12 mL) in H20 (60 mL) stirred at 0°C over 5 min. Upon addition a paste forms which is stirred for 1 h at room temperature. The white paste is sparged with N2 for 30 min and diluted with H20 (10 mL) to form a clear solution. Then S02 is bubbled through the solution for 15 min reducing its pH to 2. Upon cooling the resulting solid is filtered, rinsed with H20, and oven dried to yield 4-carbamoylnicotinic acid (7 g, 76%) as a white solid. 1H NMR (DMSO) δ 8.93 (lH,s), 8.76 (1H, d, J = 5.0 Hz), 8.08 (1H, s), 7.62 (1H, s), 7.45 (1H, d, J = 5.0 Hz).

Isoquinolinic imide. 4-Carbamoylnicotinic acid (280 mg, 1.68 mmol) is heated neat at 200°C for 5 h to yield isoquinolinic imide (177.2 mg, 71%) as a white solid. 1H NMR (DMSO) 5 11.68 (1H, s), 9.12-9.03 (2H, m), 7.80 (1H, d, J = 5.1 Hz). 3 -Amino isonicotinic acid. Bromine (1.71 g) is added to 10% KOH (30 mL) on ice. The resulting solution is added to finely ground isoquinolinic imide (1.46 g, 9.86 mmol). Upon addition the mixture begins to foam. When all of the solid is dissolved, aqueous KOH (15%, 7 mL) is added and the mixture is heated to 80°C for 1 min then cooled. The mixture is neutralized with S02, and cooled to 0°C until precipitation occurs. The solid is collected by suction filtration and washed with H20, and dried in a vacuum oven to yield of 3-amino isonicotinic acid (485 -104- mg, 36%) as a white solid. 1H NMR (DMSO) δ 9.5-8.8 (2H, brs), 8.20 (1H, s), 7.70 (IH, d, J = 5 Hz), 7.46 (1H, d, J = 5 Hz) . 3 H-P yridof 3 ,4-dlpy rimidin-4-one. A mixture of 3-amino isonicotinic acid (485 mg, 3.51 mmol) in formamide (3 mL) is heated to 160°C for 12 h. Upon cooling, the resulting solid is filtered and washed with H2O and dried in a vacuum oven to yield 3H-pyrido[3,4-d]pyrimidin-4-one (373 mg, 72%). 1H NMR (DMSO)8 12.60 (1H, brs), 9.06 (lti s), 8.68 (1H, d, J = 5.3 Hz), 8.23 (1H, s), 7.96 (1H, d, J = 5.1 Hz) . 4-Thioxopyrido[3 ,4-d]pyrimidine. Phosphorus pentasulfide (1.25 g, 2.74 mmol) is added to a solution of 3H-pyrido[3,4-d]pyrimidin-4-one (366 mg, 2.49 mmol) in pyridine (4 mL). The mixture is refluxed for 4 h under N2. The resulting black tar is dissolved in H2O, and a solid forms. The solid is filtered and washed with H2O and dried in a vacuum oven to yield 4-thioxopyrido[3,4-d]pyrimidine (369.8 mg, 91%) as a yellow solid. 1H NMR (DMSO)6 14.48 (1H,brs), 9.13 (lH,s), 8.70 (1H, d, J = 5.4 Hz), 8.29 (1H, s), 8.27 (1H, d, J = 5.4 Hz) . 4-Methylthiopyridof3,4-d]pyrimidine. A mixture of 4-thioxopyrido[3,4-d]pyrimidine (369.8 mg, 2.26 mmol), triethylamine (0.6 mL, 4.5 mmol), DMSO (2 mL), and iodomethane (0.24 mL, 3.96 mmol) is stirred under N2 at 25°C for 12 h. The mixture is poured into H2O and the resulting solid is filtered and dried in a vacuum oven to yield 4-methylthiopyrido[3,4-d]pyrimidine (222 mg, 55%) as a brown solid. 1H NMR WLC 0131 PUS -105- (DMSO) δ 9.51 (1H, s) , 9.18 (1H, s) , 8.79 (1H, d, J = 8 Hz), 7.97 (1H, d, J = 8 Hz) . 4-Anilinopyrido [3 , -dl pyrimidine A mixture of 4 -methylthiopyrido [3 , 4 -d] pyrimidine ( 75 mg, 0.42 mmol) , and aniline (1 mL) is heated to 100'C under N2 for 2 h. The reaction mixture is then chromatographed on silica using MPLC and eluting with a gradient system (CHC13 to 5% MeOH in CHC13) . The fractions are concentrated under reduced pressure, and the resulting solid is recrystallized from Et20 to yield 4-anilinopyrido [3 , 4 -d] pyrimidine (21.2 mg, 23%) as a yellow solid. lH MR (DMSO) δ 10.09 (1H, s) , 9.18 (1H, s) , 8.74 (1H, d, J = 5.3 Hz), 8.46 (1H, d, J = 5.8 Hz), 7.89 (2H, d, J = 8.5 Hz) , 7.45 (2H, t, J = 7.9 Hz), 7.21 (1H, t, J = 7.4 Hz).

Example 56 4- (3 -Bromoanilino) pyrido f3 , 4-dl pyrimidine A mixtui-e of 4 -methylthiopyrido [3 , 4- d] pyrimidine ( 75 mg, 0.42 mmol) (see previous experimental)., and 3 -bromoaniline (1 mL) is heated to 100 "C under N2 for 2 h. The reaction mixture is then chromatographed on silica using MPLC and eluting with a gradient system (CHC13 to 5% MeOH in CHC13) . The fractions are concentrated under reduced pressure, and the resulting solid is recrystallized from Et20 to yield 4- (3 -bromoanilino) pyrido [ , 4 -d] pyrimidine (66 mg, 52.7%) as a light brown solid. Ή NMR (DMSO) δ 10.15 (1H, s) , 9.21 (1H, s) , 8.80 (1H, s) , 8.76 (1H( d, J = 5.8 Hz), 8.44 (1H, d, J=5.6 Hz), 8.25 (1H, s) , 7.93 (1H, d, J = 7.7 Hz), 7.45-7.37 (2H, m) .

WLC 0131 PUS -106- Example 57 4- (3-Bromoariilino) -6-fluoropyrido Γ3.4-dl pyrimidine 5- ΪΝ- ( tert-Butoxycarbonyl) amino! -2-fluoropyridine . 5-Amino-2-fluoropyridine is prepared by hydrogenation (Pd/C) of 2-fluoro-5-nitropyridine (obtained from from 2-chloro-5-nitropyridine by reaction with KF in MeC with Ph4PBr [J.H. Clark and D.J. Macquarrie, Tetrahedron Lett., 1987, 28., 111-114] . Reaction of the crude amine with t-Boc anhydride gives 5- [N- ( tert-butoxycarbonyl) amino] -2-fluoropyridine. *H MR (CDC13 δ 8.07 (1H, s) , 8.05 (1H, m) , 6.89 (1H, dd, J = 9.2, 3.3 Hz), 6.66 (1H, m) , 1.52 (9H, s) . ' 5- IN- ( tert-Butoxycarbonyl) amino! -2-fluoropyridine-4-carboxylic acjd. Reaction of 5- [N- ( ter -butoxycarbonyl) amino] -2-fluoropyridine (5.3 g, 25 mmol) sequentially with n-BuLi and C02 as described in the following example gives 5- [N- { tert-butoxycarbonyl) amino] -2-fluoropyridine-4-carboxylic acid (1.60 g, 25%). *H NMR (DMSO) 6 9.83 (1H, brs) , 8.84 (1H, s), 7.49 (1H, d, J = 2.9 Hz), 1.47 (9H, s) . 5-Amino-2 -fluoropyridine-4-carboxylic acid . Reaction of 5- [N- ( ert-butoxycarbonyl) amino] -2-fluoropyridine-4 -carboxylic acid (1.0 g, 3.9 mmol) with TFA as described above gives 5-amino-2-fluoropyridine-4 -carboxylic acid(0.46 g, 74%). XH NMR (DMSO) δ 7.85 (1H, d, J » 1.5 Hz) , 7.23 (1H, d, J = 2.5 Hz) . 6-Fluoro-3H-pyrido f3. -dl pyrimidin-4-one Reaction of 5-amino-2-fluoropyridine-4-carboxylic WLC 0131 PUS -107- with formamide at 140 °C as above gave 6-fluoro-3H-pyrido[3,4-d]pyrimidin-4-one (-20%). XH NMR (DMSO) δ 12.48 (1H, m) , 8.74 (1H, s) , 8.16 (1H, s) , 7.63 (1H, d, J = 3 Hz) . 4- (3-Bromoanilino) -6 -fluoropyrido Γ3.4 -dl pyrimidine . Reaction of 6-fluoro-3H-pyrido [3 , 4 -d] pyrimidin-4-one (0.60 g, 3.6 mmol) with P0C13, followed by reaction of the crude 4,6-dihalo compound with 3-bromoaniline gives 4- (3-bromoanilino) -6-fluoropyrido [3, 4-d] pyrimidine (0.73 g, 63%). XH NMR (DMSO) δ 10.09 (1H, brs) , 8.96 (1H, s) , 8.75 (1H, s) , 8.25 (2H, m) , 7.90 (1H, brd, J = 6.5 Hz), 7.44 - 7.34 (2H, m) . ' Example 58 4- (3-Bromoanilino) -6-chloropyrido f3 , -dl pyrimidine 5- \N- (tert-butoxycarbonyl) amino! -2-chloropyridine . A mixture of 5-amino-2-chloropyridine (12.86 g, 0.1 mol ', di-tert-butyldicarbonate (24. Og, 0.11 raol) and Et3N (12.1 g, 1.12 mol) in CH2C12 (150 mL) is heated, under reflux for 12 h, cooled, and the precipitate is filtered off. The organic layer is washed with water, dried (CaCl2) and filtered through a short column of alumina. Removal of the solvent gives 5- [N- (tert-butoxycarbonyl) amino] -2-chloropyridine (11.9 g, 52%). :H NMR (CDC13) 6 8.31 (1H, d, J = 2.9 Hz), 7.94 (1H, dd, J » 8.6, 2.6 Hz), 7.24 (1H, d, J = 8.7 Hz), 7.15 (1H, m) , 1.51 (9 H, s) . 5- N- (tert-Butoxycarbonyl) aminol -2-chloropyridine-4 -carboxylic acid. A solution of 5- [N- (tert-butoxycarbonyl) amino] -2-chloropyridine (22.87 g, WLC 0131 PUS -108- 0.1 mol) and TMEDA (47 mL, 0.31 mol) in dry Et20 (600 mL) is cooled to -78 °C, and n-BuLi (10 M in hexanes, 30 mL, 0.3 mol) is added dropwise. The solution is allowed to warm to -10°C and is then kept at that temperature for 2 h, before being recooled to -78°C. Dry C02 is then bubbled in, and the resulting mixture is allowed to warm to 20 °C, before being quenched with water (300 mL) containing a small amount of NH40H. The resulting aqueous layer is washed with EtOAc, then acidified slowly with dilute HCl to precipitate 5- [iV- ( tert-butoxycarbonyl) amino] -2-chloropyridine-4-carboxylic acid (15.5 g, 57%). Ή NMR (DMSO) δ 10.00 (1H, s) , 9.13 (1H, s), 7.74 (1H, s) , 1.47 (9H, s) . 5-Amino-2-chloropyridine-4-carboxylic acid . A stirred suspension of 5- [N- (tert-butoxycarbonyl) amino] -2 -chloropyridine-4-carboxylic acid (1.91 g, 7 mmol) in CH2C12 (200 mL) is treated slowly with trifluoroacetic acid until homogeneous (ca. 12 mL) . The solution is stirred overnight and extracted with dilute H40H, and the aqueous layer is then acidified with dilute HCl to gave a precipitate of 5 -amino-2-,chloropyridine-4 -carboxylic acid (1.05 g, 87% yield). lH NMR (DMSO) 6 9.01 (2H, m) , 8.03 (1H, s) , 7.48 (1H, s) . 6-Chloro-3H-pyrido f3.4-dl pyrimidin-4-one . A solution of 5-amino-2-chloropyridine-4-carboxylic acid (8.1 g, 4.7 mmol) in formamide (100 mL) is stirred at 140 °C for 12 h. Dilution of the cooled mixture with water gives a precipitate of 6-chloro-3H-pyrido [3 , 4 - d]pyrimidin-4-one (7.3 g, 86% yield). XH NMR (DMSO) δ 12.73 (1H, m) , 8.90 (1H, d, J = 0.7 Hz), 8.23 (1H, s) , 7.97 (1H, d, J = 0.7 Hz) .

WLC 0131 PUS -109- 4.6 -Dichloropyrido Γ3.4 -dl pyrimidine . A stirred suspension of 6-chloropyrido [3 , 4-d] pyrimidin-4-one (1.82 g, 10 mmol) in P0C13 (10 mL) is heated under reflux until dissolved (ca. 2 h) and for a further 30 min. Excess reagent is removed under reduced pressure, and the residue is treated with a mixture of CH2C12 and ice-cold aqueous Na2C03. The resulting organic layer is dried (Na2S04) and evaporated to give a quantitative yield of crude, unstable, 4 , 6-dichloropyrido [3 , 4-d] pyrimidine, which is used directly in the next step. XH NMR (CDC13) δ 9.38 (1H, d, J = 0.5 Hz), 9.19 (1H, s) , 8.09 (1H, d, J = 0,5 Hz) . 4- (3-Bromoanilino) -6-chloropyrido Γ3.4-dl pyrimidine . A mixture of the above crude dichloropyrimidine and 3 -bromoaniline (3.8 g, 22 mmol) is dissolved in i-PrOH (100 mL) . One drop of cone. HC1 is added to initiate the reaction, and the mixture is then heated under reflux for 30 min, cooled, and diluted with watei to precipitate 4- (3-bromoanilino) -6-chloropyrido [3 , 4-d] pyrimidine (1.26 g, 38% yield). H NMR (DMSO) δ J.0.12 (1H, s) , 9.03 (1H, s) , 8.77 (1H, S) , 8.63 (1H, s) , 8.21 (1H, s) , 7.89 (1H, d, J = 8.1 Hz) , 7.43-7.32 (2H, m) .

Example 59 4- (3-Bromoanilino) -6 -methoxypyrido f3.4 -dl pyrimidine Treatment of 4- (3-bromoanilinp) -6- fluoropyrido [3 , 4-d] pyrimidine (see a previous experimental) at 100 °C in a pressure vessel with sodium methoxide in methanol gives 4- (3-bromoanilino) - 6 -methoxypyrido [3, 4-d] pyrimidine. ¾ NMR (DMSO) δ 9.93 WLC 0131 PUS -110- (1H, s) , 8.94 (1H, s) , 8.61 (1H, s) , 8.26 (1H, brs) , 7.94 (1H, brd, J" = 7.6 Hz), 7.88 (1H, s) , 7.43-7.32 (2H, m) , 4.01 (3Hf s) .

Example 60 4- (3-Bromoanilino) -6-methYlaminopyrido Γ3.4-dl yrimidine Treatment of 4- (3-bromoanilino) -6 -fluoropyrido [3 , 4-d] pyrimidine (0.20 g, 0.63 mmol) (see a previous experimental) at 100 °C in a pressure vessel with methylamine in ethanol followed by chromatography on alumina (CH2Cl2/MeOH, 99:1) gives 4- (3-bromoanilino) -6-methylaminopyrido [3, 4-d] pyrimidine ' (0.07 g, 34%). 1H NMR (DMSO) δ 9.69 (1H, s) , 8.75 (1H, s) , 8.41 (1H, s) , 8.21 (1H, brs), 7.93 (1H, brd, J = 7.6 Hz), 7.41-7.28 (2H, m) , 7.06 (1H, s) , 6.82 (1H, q, J = 5.0 Hz), 4.95 (3H, d, J = 5.0 Hz).

Example 61 4- (3-Bromoanilinoyv-6-dimethylaminopyrido [3,4- dl pyrimidine * Treatment of 4- (3-bromoanilino) -6- fluoropyrido [3 , 4-d] pyrimidine (see a previous experimental) at 100 °C in a pressure vessel with dimethylamine in ethanol gives 4- (3-bromoanilino) -6- dimethylaminopyrido [3 , 4-d] pyrimidine. ¾ NMR (DMSO) δ 9.71 (1H, s), 8.83 (1H, s) , 8.43 (1H, s) , 8.21 (1H, brs), 7.94 (1H, brd, J = 7.5 Hz), 7.42-7.29 (2H, m) , 7.26 (1H, S) , 3.17 (6H, s) .

- - Example 62 4- (Benzylainino) pyrido [3, 4-d] pyrimidine A mixture of 4 -methylt iopyrido [3, -d] yrimidine (74 mg, 0.41 mmol) (see a previous experimental), and benzylamine (1 mL) is heated to 100°C for 2 h. On cooling the mixture is concentrated under reduced pressure and purified directly by preparative tic on silica gel eluting with CH2C12, to yield 4- (benzylamino)pyridoI3, 4-d] pyrimidine (21.2 mg, 20%). XH NMR (DMSO) δ 9.21 (1H, t, J = 5.8 Hz), 9.19 (1H, s) , 8.63 (1H, d, J = 5.8 Hz ) 8.58 ,(H, s), 8.20 (IB ,d, J = 5.1 Hz), 7.41-7.30 (4H, m) , 7.26 (1H, t, J=7.1 Hz).

Example 63 4- ( 3-Bromoanilino) pyrido [2, 3-d] pyrimidine 3H-pyrido [2, 3-d]pyrimidin-4-one. 2-Amino nicotinic acid (15 g, 108.6 mmol) in formamide (35 mL) is heated to 165-170°C for 3.5 h. Upon cooling a solid precipitates. The solid is filtered and washed with H20 and dried in a vacuum oven to give 3H-pyrido[2, 3-d]pyrimidin-4-one (7.87 g, 49.4%). 1H NMR (DMSO) δ 12.50 (1H, s),8.97 (1H, dd, J = 1.9, 4.5 Hz), 8.53 (1H, dd, J = 2.1, 7.9 Hz), 8.34 (1H, s), 7.57 (1H, dd, J = 4.6, 8.0 Hz) . 4-Thioxopyrido [2, 3-d] pyrimidine . Phosphorus pentasulfide (6 g, 13.5 mmol) is added to a solution of 3H-pyrido [2, 3-d]pyrimidin-4-one (2 g, 13.5 mmol) in pyridine (50 mL) . The mixture is refluxed for 3 h. Upon cooling a solid formed and the pyridine is decanted off. The solid is suspended in H20 (20 mL) -112- and then filtered and dried in a vacuum oven to yield 4-thioxopyrido [2, 3-d] pyrimidine (1.72 g, 78%). :Η MR (DMS0)8 9.06 (1H, dd, J = 1.9, 4.3 Hz), 8.90 (1H, dd, J = 1.9, 8.2 Hz), 8.36 (1 H, s) , 7.65 (1H, dd, J = 4.3, 8.2 Hz) . 4-Methylthiopyrido [2, 3-d] pyrimidine . A mixture of 4-thioxopyrido [2, 3-d] pyrimidine (100 mg, 0.76 mmol), triethylamine (154 mg, 1.52 mmol), DMSO (2 rnL) , and iodomethane (161 mg, 1.14 mmol) is stirred for 12 h at 25°C. The mixture is poured into H20 and extracted with EtOAc. The combined extracts are washed with water, saturated brine, and dried (MgSO^), and the solvent is removed under reduced pressure to yield 4-methylthiopyrido [2, 3-d] pyrimidine (134 mg, quant.). 1H NMR (DMSO) δ 9.25 (1H, dd, J = 1.8, 4.2 Hz), 9.17 (1H, s), 8.59 (1H, dd, J = 1.9, 8.2 Hz), 7.75 (1H, dd, J = 4.3, 8.2 Hz), 2.73 (3H, s).

A mixture of 4-methylthiopyrido [2, 3-d] yrimidine (157 mg, 0.89 mmol, and 3-bromoaniline (1 mL) is heated to 100°C for 2 h. On cooling a precipitate forms which is filtered then washed with EtOH and air dried to yield 4- ( 3-bromoan.ilino) pyrido [2, 3-d] pyriraidine (55.5 mg, 20%. XH NMR (DMSO) δ 10.13 (1H, s) , 9.11 (1H, dd, J = 1.7, 4.3 Hz), 9.01 (1H, dd, J = 1.7, 8.2 Hz), 8.81 (1H, s), 8.22 (1H, s), 7.90 (1H, d, J = 7.7 Hz), 7.71 (1H, dd, J = 4.3, 8.0 Hz) , 7.40 (2H,m).

Example 64 4- (3-Bromoanilino) -7-fluoropyrido [2, 3-d] pyrimidine -113- 2, 6-Difluoronicotinic acid. 2, 6-Difluoropyridine (7.89 mL, 0.087 mmol) is added dropwise under N2 at 78°C to a stirred solution of lithium diisopropylamide (59.0 mL of a 1.5 N solution in cyclohexane, 0.089 mmol) in THF (250 mL) . After 2 h at 78°C, a stream of dry C02 is passed through the solution and the mixture is diluted with water and washed with EtOAc. The aqueous portion is neutralized with 3 N HC1, extracted with EtOAc and worked up to give 2, 6-difluoronicotinic acid (13.4 g, 97%). XH NMR (DMSO)6 8.59 (1H, dd, J = 9.2, 8.2 Hz), 7.30 (1H, dd, J = 8.2, 2.1 Hz), 4.03 (1H, brs). 2, 6-Difluoronicotinamide. A solution of 2, 6-difluoronicotinic acid (7.4 g, 0.046 mmol) and S0C12 (20 mL) in 1 , 2-dichloroethane (60 mL) containing DMF (1 drop) is heated under reflux for 4 h, then concentrated to dryness under reduced pressure. The residue is dissolved in Et20 (100 mL) , cooled to 0°C, and treated dropwise with concentrated ammonia (10.0 mL, 0.17 mmol). After 10 min the solution is washed with aqueous NaHC03 and worked up to give 2, 6-difluoronicotinamide (5.61 g, 76%). H NMR (CDC13) δ 8.70 (1H, dd, J = 9.6, 8.3 Hz), 7.00 ( lH.ddd, J = 8.3, 2.9, 1.1 Hz), 6.71, 6.55 (1H, 1H, 2 brs). 2-Amino-6-fluoronicotinamide . A solution of 2, 6-difluoronicotinamide (4.68 g, 0.029 mmol) in dry formamide (30 mL) is saturated with ammonia and allowed to stand at room temperature for 24 h. Water (50 mL) is added and the resultant precipitate is filtered off and washed well with water, to give 6-amino-2-fluoronicotinamide (1.41 g, 31%) mp 236-237°C. XH NMR (DMSO) δ 7.89 (1H, dd, J = 10.4, 8.4 Hz) , -1 14- 7.31, 7.16 (1H, 1H, 2 brs,), 6.93 (2H, brs), 6.36 (1H, dd, J - 8.4, 2.4 Hz).

The filtrate and washings are combined and extracted exhaustively with EtOAc, and the extract is chromatographed on silica gel. EtO Ac/petroleum ether (1 : 1) elutes forerun, while EtO Ac/petroleum ether (2: 1) and then EtOAc gives 2-amino-6-fluoronicotinamide (1.57 g, 35%), mp (EtO Ac/petroleum ether) 199-200°C [Rogers, R.B. et al, U.S. patent no. 4,383,851, record mp 198-200°C]. 1H NMR (DMSO) δ 8.13 (1H, dd, J = 10.4, 8.4 Hz), 7.90, 7.30 (1H, 1H, 2 brs), 7.65 (2H, brs), 6.23 (1H, dd, J = 8.4, 2.6 Hz).

A suspension of 2-amino-6-fluoronicotinamide (0.74 g, 4.77 mmol) in triethyl orthoformate (25 mL) is heated at reflux for 8 h. After cooling to room temperature the precipitate is filtered off and washed well with petroleum ether to give 7-fluoropyrido[2,3-d]pyrimidin-4(3H)-one (0.76 g, 96%). 1H NMR (DMSO) δ 12.75 (1H, brs), 8.66 (1H, dd, J = 10.4, 8.4 Hz), 8.38 (1H, s), 7.33 (1H, dd, J = 8.4, 2.6 Hz). 4-(3-BromoanilinoV7-fluoropyrido[2.3-d] pyrimidine. A suspension of 7-fluoropyrido[2,3-d] pyrimidin-4(3H)-one (0.20 g, 1.21 mmol) in POCl3 (10 mL) is heated under reflux for 2 h. The volatiles are then removed under reduced pressure, and the residue is partitioned between aqueous NaHC03 and EtOAc. The organic extract is worked up to give crude 4-chloro-7-fluoropyrido[2,3-d]pyrimidine, which is used directly in the next reaction. A solution of this product (0.20 g, 1.09 mmol) and 3-bromoaniline (0.23 mL, 2.18 mmol) in propan-2-ol (1.0 mL) and THF (10 mL) containing a WLC 0131 PUS -115- trace of cone. HCl is stirred at 20 °C for 1 h, and then concentrated to dryness. The residue is dissolved in EtOAc, washed with aqueous NaHC03, and worked up to give an oil, which is chromatographed on silica gel. Elution with EtOAc/petroleum ether (1:5) gives 3-bromoaniline, while EtoAc/petroleum ether (1:1) elutes 4- (3-bromoanilino) -7-fluoropyrido [2 , 3 -d]pyrimidine (0.18 g, 47%), mp (MeOH) 211-213 °C. XH NMR (DMSO) δ 10.18 (1H, brs) , 9.17 (1H, t, J = 8.6 Hz), 8.80 (1H, s) , 8.17 (1H, t, J = 1.8 Hz), 7.85 (1H, dt, Example 65 7 -Amino-4- (3-bromoanilino) pyrido [2.3-dl pyrimidine A solution of 4- (3-bromoanilino) -7- fluoropyrido [2, 3-d] pyrimidine (0.20 g, 0.63 mmol) in EtOH (20 mL) is saturated with ammonia and warmed at 100 °C in a pressure vessel for 30 h. The solvent is removed under reduced pressure to give 7-amino-4- (3-bromoanilino)pyrido [2, 3-d] yrimidine (0.18 g, 90%). XH NMR (DMSO) δ -9.97 (1H, brs) , 8.59 (1H, s) , 8.51 (1H, d, J = 9.3 Hz), 8.11 (1H, si brs) , 7.77 (1H, brd, J = 6.3 Hz), 7.44 (2H, brs), 7.37-7.30 (2H, m) , 6.81 (1H, d, J = 9.3 Hz).

Example 66 4- (3-Bromoanilino) -7-methYlaminopyrido Γ2.3- dl pyrimidine A solution of 4- (3-bromoanilino) -7- fluoropyrido [2 , 3-d] yrimidine (see a previous experimental) (0.20 g, 0.63 mmol), methylamine WLC 0131 PUS -116- hydrochloride (0.13 g, 1.88 mmol) and Et3N (0.30 mL) 2.19 mmol) in EtOH (15 mL) is heated at 100 °C in a pressure vessel for 18 h. The solvent is removed under reduced pressure, and the residue is partitioned between EtOAc and water. Workup of the organic layer gives 4- (3-bromoanilino) -7- (methylamino) yrido [2 , 3 -d] pyrimidine (0.16 g, 77%). ¾ NMR (DMSO) δ 9.53 (1H, S) , 8.54 (1H, S) , 8.41 (1H, d, J = 8.1 Hz) , 8.17 (1H, t, J = 1.8 Hz), 7.83 (1H, dd, J = 8.0, 1.9 Hz), 7.66 (1H, brs) , 7.32 (1H, t, J = 8.0 Hz), 7.24 (1H, dd, J = 8.0, 1.8 Hz), 6.77 (1H, d, J - 8.1 Hz), 2.92 (3H, d, J = 4.8 Hz) .

Example 67 4- (3-Bromoanilino) -7-dimethylaminopyrido Γ2.3-dl pyrimidine Reaction of 4- (3-bromoanilino) -7- fluoropyrido [2 , 3-d] pyrimidine (see a previous experimental) (0.12 g, 0.38 mmol) with dimethylamine hydrochloride (92' mg, 1.13 mmol) and Et3N (0.18 mL, 1.32 mmol) in EtOH (15 mL) at 100 °C for 18 h in a pressure vessel, followed by evaporation of the solvent and workup, gives 4- (3-bromoanilino) -7- (dimethylamino) pyrido [2, 3-d] pyrimidine (0.11 g, 84%) . XH NMR (DMSO) δ .9.58 (1H, brs), 8.56 (1H, d, J = 9.3 Hz), 8.54 (1H, s) , 8.18 (1H, t, J" = 1.9 Hz), 7.84 (dt, Jd = 8.0, Hz, Jc = 1.9 Hz), 7.33 (1H, dd, J = 8.1, 8.0 Hz) 7.25 (1H, dt, Jd = 9.3, Hz, Jt = 1.9 Hz), 7.10 (1H, d, J = 9.3 Hz) , 3.18 (6H, s) .

LC 0131 PUS -117- Example 68 4- (3-Bromoanilino) -7-methoxypyrido [2 , 3-dl pyrimidine A solution of 4- (3-bromoanilino) -7-fluoropyrido [2, 3-d] pyrimidine (0.26 g, 0.81 mmol) and sodium methoxide (prepared from 75 mg of sodium, 3.26 mmol) in dry eOH (15 mL) is heated at 90 °C in a pressure vessel for 18 h. The mixture is poured into water and extracted with EtOAc to give 4- (3-bromoanilino) - 7-methoxypyrido [2, 3-d] pyrimidine (0.23 g, 86%). ¾ MR (DMSO) 6 9.88 (1H, brs) , 8.82 (1H, d, J = 8.9 Hz), 8.71 (1H, s) , 8.18 (1H, dd, J = 8.0, 1,9 Hz), 7.36 (1H, dd, J = 8.1, 8.0 Hz) , 7.29 (1H, ddd, J = 8.1, 1.9, 1,9 Hz) 7.15 (1H, d, J = 8.9 -Hz), 4.01 (3H, s) .

Example 69 4-Benzylamino-7-methylaminopyrimido Γ4 , 5-dl pyrimidine S-Ethylisothiouronium iodide. A solution of thiourea (3.80 g, '<50 mmol) and iodoethane (4 mL, 50 mmol) in MeOH (100 mL) is refluxed for 24 h. The solvent is st-ripped under reduced pressure, and the residual light yellow oil, is dried under vacuum, solidifying spontaneously. The desired compound (13.98 g) is obtained quantitatively.

-Amino- 5-cyano-2-ethylthiopyrimidine A' suspension of NaOMe (2.7 g, 50 mmol) in EtOH (200 mL) is added to a mixture of S-ethylisothiourea hydroiodide (11.58 g, 50 mmol), ethoxymethylenemalo- nonitrile (6.1 g, 50 mmol) and ethanol (250 mL) at 25 °C. The reaction mixture is refluxed under N2 for 2 h, and then the solution is concentrated on a hot WLC 0131 PUS -118- plate until precipitation is observed. After cooling, the solid is collected by suction filtration and is stirred in water at 25°C. Filtration and vacuum oven drying affords 4-amino-5-cyano-2-ethylthiopyrimidine (4.02 g, 45%) as a brown solid. 1H MR δ (DMSO) 8.45 (1H, s) , 7.90 (2H, brs) , 3.00 (2H, q, J = 7.3 Hz), 1.27 (3H, t, J = 7.3 Hz) . 4 -Amino-2 -ethylthiopyrimidine-5-carboxamide . 4-Amino-5-cyano-2-ethylthiopyrimidine (4.0 g, 22.3 mmol) is added to sulfuric acid (cone, 4.3 ttiL) in small portions. The mixture is then stirred under N2 at 40°C for 1.5 h. The reaction is quenched with ice-water and H40H is used to adjust the pH to -9. The solid is collected via suction filtration and dried in a vacuum oven overnight. 4-Amino-2-ethylthiopyrimidine- 5 -carboxamide (2.58 g, 58%) is obtained as a light brown solid. 1H NMR: (DMSO) δ 8.52 (1H, s) , 7.98 (2H, brs), 7.42 (2H, brs), 3.04 (2H, q, J = 7.3 Hz), 1.27 (3H, t, J - 7.3 Hz). 4-0xo-7-ethylthio-3H-pyrimido Γ4.5-dl pyrimidine .. A mixture of 4 -amino-2 -ethylthiopyrimidine- 5-carboxamide (4.66 g, 23.5 mmol) and triethyl orthoformate (150 mL) is refluxed under N2 for 24 h, and is then cooled to 25°C. The brown solid is isolated by suction filtration and dried in a vacuum oven to give 4-oxo-7-ethylthio-3H-pyrimido [4 , 5-d] pyrimidine ( 3.54 g, 72%). ¾ NMR: (DMSO) δ 12.80 (1H, s), 9.20 (1H, s) , 8.45 (1H, s) , 3.18 (2H, q, J = 7.4 Hz), 1.35 (3H, , J = 7.4 Hz). 4-Thiono-7-ethylthio-3H-pyrimido Γ4.5-dl pyrimidine . A mixture of 4 -oxo-7-ethylthio-3H- WLC 0131 PUS -119- pyrimido [4 , 5 -d] yrimidine (1.33 g, 6.7 mmol) , P2S5 (1.48 g, 6.6 mmol) and pyridine (15 mL) is refluxed under N2 for 3 h. The pyridine is then stripped under reduced pressure, and the residue is dissolved in NaOH solution (0.5 M, 75 mL) and boiled with charcoal.

After filtration, the filtrate is neutralized with acetic acid to generate a gold brown solid. Buchner filtration and drying in a vacuum oven affords 4-thiono-7-ethylthio-3H-pyrimido [4, 5-d] pyrimidine (1.42g, 95%). 1H NMR (DMSO) δ 9.47 (1H, s) , 8.46 (1H, S) , 3.20 (2H, q, J = 7.3 Hz) , 1.35 (3H, t, J = 7.3 Hz) . 7-Ethylthio-4-methylthiopyrimido Γ4.5-dl pyrimidine . The same procedure described for 7-amino-4-methylthiopyrido [4 , 3-d] pyrimidine in Example 21 is used. lH NMR (DMSO) δ 9.52 (1H, s) , 9.15 (1H, s) , 3.23 (2H, q, J = 7.3 Hz), 2.72 (3H, s) , 1.38 (3H, t, J = 7.3 Hz) . 4-Benzylamino-7-ethylthiopyrimido Γ4 , 5- dl yrimidine . The same procedure described for 7- amino-4-anilijiopyrido [4 , 3-d] pyrimidine in example 21 is used. 4-Benzylamino-7-methylaminopyrimido Γ4.5- dl pyrimidine . 4-Benzylamino-7-ethylthiopyrimido [4,5- d] pyrimidine in EtOH containing excess methylamine is heated to 150°C in a stainless steel bomb for 5h. The solid is filtered off and dried to give 4-benzylamino- 7-methylaminopyrimido [4 , 5-d] yrimidine .

WLC 0131 PUS -120- Example 70 4-Benzylamino-7-hvdrazinopyrimido Γ4 , 5-dl pyrimidine 4 -Benzylamino-7-ethylthio pyrimido [4,5-d] pyrimidine in EtOH containing excess hydrazine is heated to 150°C in a stainless steel bomb for 5h. The solid is filtered off and dried to give 4-benzylamino-7-hydrazinopyrimido [4 , 5-d] pyrimidine.

Example 71 4- (3 -Bromoanilino) thieno [3.2-dl pyrimidine hydrochloride 3H-Thieno f3.2-dl pyrimid-4-one . A mixture of methyl 3-aminothi0phene-2-carboxylate (1 g, 6.3 mmol) and formamide (2 g) is heated at 240 'C for 10 min.

Upon cooling,a precipitate appeared. It is dissolved in EtOH and filtered. The filtrate is concentrated under reduced pressure and the residue is purified by silica gel chromatography eluting with 10% MeOH in CH2C12 to yield 3H-thieno [3 , 2-d] pyrimid-4-one (249 mg, 26%) as a solid. lH NMR (DMSO) δ 12.61 (1H, brs) , 8.20 (1H, s) , 8.17 (1H, d, J = 5 Hz) , 7.42 (1H, d, J = 5 Hz) . 4-Chlorothieno Γ3 , 2 -dT pyrimidine . To a solution of DMF (170.3 μΙ>, 2.2 mmol) and dichloroethane (1.2 mL) at O'C under N2>, oxalyl chloride (279.2 mg, 3.2 mmol) is added slowly and stirred for 10 min.

WLC 0131 PUS -121- 3H-thieno [3 , 2-d] yrimid-4-one (152.2 mg, 1.0 mmol) is added and refluxed for 5 h. The reaction mixture is poured into water and extracted with CH2C12. The organic layer is stripped under reduced pressure to yield 4 -chlorothieno [3 , 2-d] pyrimidine (140 mg, 82%) as a yellow solid. Ή NMR (DMSO) δ 9.05 (1H, s) , 8.62 (1H, d, J = 5 Hz) , 7.79 (1H, d, J = 5 Hz) . 4- (3-Bromoanilino) thieno f3 , 2-dl pyrimidine hydrochloride . A mixture of 4 -chlorothieno [3 , 2-d] -pyrimidine (135 mg, 0.79 mmol) and 3-bromoaniline (95 L, 0.89 mmol) in 2-methoxyethanol (2 mL) is heated to 79 'C for 30 min. The resulting precipitate is fil-tered and washed with CH2C12 to yield 4-(3-bromo-anilino) thieno [3 , 2-d] pyrimidine hydrochloride (195.5 mg, 72%) as a light yellow solid. XH NMR (DMSO) δ 11.33 (1H, s) , 8.94 (1H, s) , 8.23 (1H, s) , 8.53 (1H, d, J = 5.3 Hz), 8.07 (1H, s) , 7.77 (1H, d, J = 7.9 Hz), 7.6 (1H, d, J = 5.3 Hz), 7.48 (2H, m) .

Example 72 4 -Benzylaminothieno ί3.2 -dl pyrimidine As described in the previous experiment 4 - chlorothieno [3 , 2-d] pyrimidine (100 mg, 0.586 mmol) and benzylamine (710 μΐ-., 0.645 mmol) in 2-methoxyethanol (2 mL) yields 4 -benzylaminothieno [3 , 2 - d]pyrimidine (37 mg, 26%). ¾ NMR (DMSO) δ 8.42 (1H, s) , 8.12 (1H, d, J = 5.5 Hz), 7.39 (1H, d, J = 5.3 Hz), 7.40-7.30 (4H, m) , 7.24 (1H, t, J = 6.8 Hz). -122- Example 73 4-(3-Bromoanilino thienor2,3-dlpyrimidine Methyl 2-aminothiophene-3-carboxylate. A mixture of methyl cyanoacetate (3.25 g, 32.3 mmol), 1,4 dithiane-2,5 diol (5 g, 32.8 mmol), triethylamine (1 mL, 7.71 mmol) in EtOH (50 mL) is stirred at 40°C for 1 h. The cooled solution is eluted through a silica plug with CH2C12. The filtrate is stripped to dryness to give crude methyl 2-aminothiophene-3-carboxylate which is carried on to the next reaction. Ή NMR (DMSO) δ 7.26 (1H, s), 6.82 (1H, d, J = 5.8 Hz), 6.28 (1H, d, J = 5.8 Hz), 3.69 (3H, s) . 3H-Thienof2,3-d pyrirnidin-4-one. A solution of methyl 2-aminothiophene-3-carboxylate (602.1 mg, 3.83 mmol) in formamide (5 mL) is heated at 200°C for 12 h. The resulting tar is dissolved in CH2C12 (lOmL) then placed on a silica plug and eluted with 10% MeOH in 0Η2Ο2. The filtrate is stripped under reduced pressure and the resulting solid is washed with EtOH to yield 3H-thieno[2,3-d]pyrimidin-4-one (231.4 mg, 40%) as an orange solid. Ή NMR (DMSO) δ 12.50 (1H, brs), 8.13 (1H, s), 7.60 (lH, d, J = 5.8 Hz), 7.41 (1H, d, J = 6.0 Hz) . 4-Chlorothienof2,3-d1pyrimidme. To a solution of DMF (90 μL) and CH2C12 (2 mL) at 0°C under N2, oxalyl chloride (148 mg, 1.2 mmol) is added slowly and stirred for 10 min. 3H-Thieno[2,3-d]pyrimidin-4-one (81 mg, 0.52 mmol) is added as a solid to the solution and warmed with a heat gun until the solid dissolves. The reaction is stirred at 25°C for 12 h under N2. The reaction mixture is poured into water LC 0131 PUS -123- and extracted with CH2C12 . The phases are separated and the organic layer is dried (Na2S04) and stripped under reduced pressure to yield 4-chlorothieno [2 , 3 -d]pyrimidine (87.6 mg, 97%) as a solid. 1H NMR (DMSO) δ 8.96 (1H, s) , 8.17 (1H, d, J = 6.0 Hz), 7.62 (1H, d, J = 6.0 Hz) . 4- (3-Bromoanilino) thieno Γ2.3-dT pyrimidine hydrochloride . A mixture of 4-chlorothieno [2, 3-d] pyrimidine (135 mg, 0.79 mmol) and 3-bromoaniline (95 μΐ.., 0.89 mmol) in 2-methoxyethanol (2 mL) is heated to 79 *C for 30 min with stirring. The resulting solid is filtered and washed with CH2C12 to yield 4- (3 -bromoanilino) thieno [2, 3-d] pyrimidine hydrochloride (197 mg, 73%). ¾ NMR (DMSO) δ 9.99 (1H, s) , 8.60 (1H, s) , 8.23 (1H, s) , 7.98 (1H, d, J = 6.0 Hz), 7.88 (1H, d, J =8.0 Hz) , 7.79 (1H, d, J = 6.0 Hz), 7.37 (1H, t, J = 8.0 Hz), 7.30 (1H, d, J = 8.0 Hz) .

Example 74 4 -Benzylaminopyrrolo Γ2.3 -dl pyrimidine 4 -Benzylaminopyrrolo [2 , 3 -d] pyrimidine is prepared as described previously. G.H. Hitchings, K.W. Ledig and R. A. West, U.S. Patent No. 3,037,980, 1962; Chemical Abstracts 1962, 57, 15130c.

Example 75 N6- (3-Bromophenyl) adenine A mixture of 6-chloropurine (1.0 g, 6.47 mmol), 3-bromoaniline (0.78 mL, 7.12 mmol), and cone HC1 (4 drops) in isopropanol (10 mL) is stirred at WLC 0131 PUS -124- 80 *C for 5 h. Upon cooling, it precipitates. The solid is filtered and washed with isopropanol and air dried to yield N6- (3 -bromophenyl) adenine (1.93 g, 91%) as a light yellow solid. lH NMR (DMSO) δ 11.38 (1H, s) , 8.78 (1H, s), 8.75 (1H, s) , 7.90 (1H, d, J = 8.0 Hz) , 7.38-7.34 (2H, m) .

Example 76 N6 -Benzyladenine N6-Benzyladenine is available commercially from the Aldrich Chemical Company, 1001 West Saint Paul Avenue, Milwaukee, Wisconsin 53233.

Example 77 7 -Amino-4- (3 -^methylanilino) pyrido Γ4.3-dl pyrimidine A mixture of 7-amino-4-methylthiopyrido [4 , 3 - d]pyrimidine (217 mg, 1.13 mmol) and m-toluidine (1.50 g, 14.0 mmol) is stirred at 155 °C for 30 min. The resulting product is chromatographed over silica gel (5% MeOH/CH2Cl2) to give 7-amino-4- (3- methylanilino) pyrido [4, 3-d] pyrimidine (190 mg, 67%) as a pale yellow solid. XH NMR (DMSO) δ 9.81 (1H, brs) , 9.34 (1H, s) , 8.38 (1H, s), 7.60 (2H, s) , 7.26 (1H, dd, J = 8.5, 7.6 Hz) , 6.95 (1H, d, J = 7.4 Hz) , 6.63 (2H, brs), 6.44 (1H, s) , 2.33 (3H, s) . -125- Example 78 7-Amino-4-(4-methoxyanilino)pyrido 4,3-d1pyrimidine A mixture of 7-amino-4-methylthiopyrido[4,3-d]pyrimidine (129 mg, 0.62 mmol) and 4-methoxyaniline (0.15 g, 1.2 mmol) in ethanol (5 mL) was heated at 40°C for 16 h, and then refluxed for 3 h. The reaction mixture was cooled to 0°C overnignt, and the solid was collected by vacuum filtration and recrystallized from isopropanol to give 7-amino-4-(4-methoxyariilino)pyrido[4,3-d]pyrimidine (42 mg, 25%) as a yellow solid. 1H NMR (DMSO) δ 10. oo (lH, brs), 9.31 (1H, s), 8.35 (1H, s), 7.62 (2H, d, J 9 . 2 = Hz), 6.96 (2H, d, J = 9.2 Hz), 6.70 (2H, slbrs), 6.41 (lH, s), 3.77 (3H, s).

Example 79 4- 3-BromoanilinoV6-(piperidin-t-yl)p idor3^-d]pyrimidine Treatment of 4-(3-bromoanilino)-6-fluoropyrido [3,4-d]pyrimidine (see a previous experimental) at 100°C in a pressure vessel with piperidine in ethanol gives 4-(3-bromoanilino)-6-(piperidin-l-yl)pyrido[3,4-d]pyrimidine.

The pharmaceutical compositions of the invention can take any of a wide variety of oral and parenteral dosage forms. The dosage forms comprise as the active components an inhibitor as defined previously.

For preparing pharmaceutical compositions, one uses inert, pharmaceutically acceptable carriers -126- that can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture with the finely divided active compounds. In the tablet, the active compounds are mixed with carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain from 5% or 10% to about 70% of active ingredients. Suitable solid carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the active compounds with encapsulating materials as carrier, providing a capsule in which the active components (with or without other carriers) are surrounded by carrier, which are thus in association with it. Similarly, cachets are included. Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral administration.

Liquid form preparations include solutions, suspensions, and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection. Liquid preparations can also be formulated in solution in aqueous polyethylene glycol solution. Aqueous solutions suitable for oral 112249/2 WLC 0131 PUS -127- use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing, and thickening agents as desired.

Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active components in water with viscous material, i.e., natural or synthetic gums, resiris, methyl cellulose, sodium carboxy^nethyl cellulose, and other well-know suspending agents.

Preferably, the pharmaceutical preparation is in unit dosage form. In such form, the preparation may be subdivided into unit doses containing ' appropriate quantities of inhibitor and other anticancer materials individually or as a combination, i.e., in a mixture . The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, for example, packeted tablets, capsules, and powders in vials or ampoules. The unit dosage farm can also be a capsule, cachet, or tablet itself or it can be the appropriate number of any of these ^in packaged form. Additionally, the unit dosage form may be a dividable form having an inhibitor in one part and other anti-cancer materials in the other part, such as, a dividable capsule, a dividable package, or a two-part ampoule, vial or the like.

The quantity of an inhibitor in unit dosages of preparation may be varied or adjusted from about 0.01 mg/kg to 100.0 mg/kg, preferably 0.03 mg/kg to less than 1.0 mg/kg of inhibitor.

WLC 0131 PUS -128- The pharmaceutical compositions preferably are constituted so that they can be administered parenterally or orally. Solutions of the active compounds as free bases and free acids or pharmaceutically acceptable salts can be prepared in water suitable mixed with a surfactant such as hydroxypropylcellulosg. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms .

The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of the microorganisms such as bacteria and fungi . The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like) , suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, WLC 0131 PUS -129- paragens, chlorobutanol , phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferred to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions of agents delaying absorption, for example, gelatin.

Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients, into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of the sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze-drying technique which yields a powder of active ingredients plus an additional desired ingredient from a previously sterile-filtered solution thereof.

As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated.

Supplementary active ingredients can also be incorporated into the compositions.

WLC 0131 PUS -130- It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suitable as unitary dosages for the mammalian subjects to be treated?" each unit containing a predetermined quantity of active materials calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the novel dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active materials and the particular therapeutic effect to be' achieved, and (b) the limitation inherent in the art of compounding such active materials for the treatment of disease in living subjects having a diseased condition in which bodily health is impaired as herein disclosed in detail.

The principal active ingredients are compounded for convenient and effective administration in effective, amounts with a suitable pharmaceutically acceptable carrier in dosage unit form as hereinbefore disclosed. A unit parenteral dosage form can, for example, contain the principal active compound, i.e. an inhibitor, in amounts ranging from about 0.5 to about 100 mg, with from about 0.1 to 50 mg being preferred. The daily parenteral doses for mammalian subjects to be treated ranges from 0.01 mg/kg to 10 mg/kg of the inhibitor. The preferred daily dosage range is 0.1 mg/kg to 1.0 mg/kg.

For oral dosages, the daily amount may range from 0.01 mg of active compound/kg of mammalian 112249/2 WLC 0131 PUS -131- subject to 100 mg/kg, preferably 0.1 to 10 mg/kg of subject .

The inhibitor described above may form commonly known, pharmaceutically acceptable salts such as alkali metal and other common basic salts or acid addition salts, etc' ■-References to the base substances are therefore intended to include those common salts known to be substantially equivalent to the parent compound and hydrates thereof.

The active compounds described herein are capable of further forming both pharmaceutically acceptable acid addition and/or base salts. All of these forms are within the scope of the present invention.

Pharmaceutically acceptable acid addition salts of the active compounds include salts derived from nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, hydrofluoric, phosphorous, and the like, as well as the salts derived from nontoxic organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, WLC 0131 PUS -132- dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, and the like.

Also contemplated are salts of amino acids such as arginate and the like and gluconate, galacturonate (see, for example, Berge, S.M. et al, "Pharmaceutical Salts", JOURNAL OF PHARMACEUTICAL SCIENCE, 66, pp. 1-19 (1977)) .

The acid addition salts of said basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner.

Preferably, an active compound can be converted to an acidic salt by treating with an aqueous solution of the desired acid, such that the resulting pH is less than 4. The solution can be passed through a C18 cartridge to absorb the compound, washed with copious amounts of water, the compound eluted with a polar organic solvent such as, for example, methanol, acetonitrile, and the like, and isolated by concentrating under reduced pressure followed by lyophilization. The free base form may be regenerated by contacting the salt form with a base and isolating the free base in the conventional manner. The free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free base for purposes of the present invention.

Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Examples WLC 0131 PUS -133- of metals used as cations are sodium, potassium, magnesium, calcium, and the like. Examples of suitable amines are Ν,Ν' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine (see, for -example, Berge, S.M. et al, "Pharmaceutical Salts-"-, JOURNAL OF PHARMACEUTICAL SCIENCE, 66, pp. 1-19 (1977) ) .

The base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner.

Preferably, an active compound can be converted to a base salt by treating with an aqueous solution of the desired base, such that the resulting pH is greater than 9. The solution can be passed through a C18 cartridge to absorb the compound, washed with copious amounts of water, the compound eluted with a polar organic solvent such as, for example, methanol, acetonitrile and the like, and isolated by concentrating under reduced pressure followed by lyophilization. The free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the conventional manner. The free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acids for purposes of the present invention.

Certain of the compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general.

WLC 0131 PUS -134- the solvated forms, including hydrated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.

Certain of the compounds of the present invention possess one or more chiral centers and such center may exist in the R(D) or S(L) configuration. The present invention includes all enantiomeric and epimeric forms as well as the appropriate mixtures thereof .

While the forms of the invention herein constitute presently preferred embodiments, many others are possible. It is not intended herein to mention all of the possible equivalent forms or ramifications of the invention. It is understood that the terms used herein are merely descriptive rather than limiting and that various changes may be made without departing from the spirit or scope of the invention.

Scheme 1. Synthesis of Preferred Groups 1-5: R- or reduc Scheme 2. Synthesis of Preferred Groups 1-5: R~ = H Scheme 3. Synthesis of Preferred Groups 6 and 8-10: R- 4. R44Sn Pd cat.

Scheme 4. Synthesis of Preferred Group 7.

Scheme 5. Synthesis of Preferred Groups 8 & 9: R3 = OR.

Scheme 6. Preferred Group 11.

Scheme 7. Synthesis of Preferred Groups 12-16. 1. KF MeCN Scheme 9. Synthesis of Preferred Groups 22-26: R- Scheme 10. Synthesis of Preferred Groups 22-26: R- = H Scheme 11. Synthesis of Preferred Groups 27 & 29-31: R- Scheme 12. Synthesis of Preferred Groups 28 Scheme 13. Synthesis of Preferred Groups 29 & 30: R- = OR Scheme 14. Preferred Group 32.

Scheme 15. Synthesis of Preferred Groups 33-36. 3. Ar(CH2)nNH2 Scheme 16. Synthesis of Preferred Groups 37-40 Scheme 17. Synthesis of Preferred Group 41: Γ3,2-α ring fusion Scheme 18. Synthesis of Preferred Group 41: T2.3-dl ring fusion Scheme 19. Synthesis of Preferred Group 42; T3.2-dl ring fusion 4. (alkylate) Scheme 20. Synthesis of Preferred Group 42: T2,3-d1 ring fusion Scheme 21. Synthesis of Preferred Group 43: T2.3-dl ring fusion.

Scheme 22. Synthesis of Preferred Group 43: T3.2-dl ring fusion.

H SH 1. PPSA N 2. (EtO)3CH/Ac20 3. MeSNa Ν0 -0Ν . (alkylate) NH2 Scheme 23. Synthesis of Preferred Group 44: T5.4-d1 ring fusion.

Scheme 24. Synthesis of Preferred Group 44: T4.5-dl ring fusion.

Scheme 25. Synthesis of Preferred Group 45: i5.4-d1 ring fusion. 4. (modify) Scheme 26. Synthesis of Preferred Group 45: F4.5-dl ring fusion.

Scheme 27. Synthesis of Preferred Group 46.

Scheme 28. Synthesis of Preferred Group 47: T5.4-dl ring fusion.

Scheme 29. Synthesis of Preferred Group 47: F4.5-dl ring fusion. 4. NBS/ NH3 Scheme 30. Synthesis of Preferred Group 48: T5.4-d1 ring fusion.

Scheme 31. Synthesis of Preferred Group 48: Γ4.5-ά1 ring fusion.

Scheme 32. Synthesis of Preferred Group 49: T3.4-d1 ring fusion Scheme 33. Synthesis of Preferred Group 49: T4.3-dl ring fusion.

Claims (1)

1. 12249/4 -145- 1. A pharmaceutical composition adapted for administration as an inhibitor of the epidermal growth factor receptor family of .tyrosine kinases, comprising a therapeutically effective amount of a compound of Formula I or a pharmaceutical salt or hydrate thereof, in admixture with a pharmaceutically acceptable excipient, diluent or carrier: Formula. I where : - 146 - 112249/ X = o, s or NH; n = 0, 1, 2; H1 = H or alkyl (1-4 carbon atoms);- if n = 2, R1 can be independently H or alkyl (1-4 carbon atoms) on either linking carbon atom; R2 is alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms), halo (fluoro, chloro, bromo, iodo) , perfluoroalkyl (1-4 carbon atoms), hydroxy, acyloxy of the formula -O-C(0)R where R is alkyl of 1-carbon atoms or cycloalkyl of 3-8 carbon atoms, amino, mono or dialkylami.no (1-4 carbon atoms in each alkyl group) , mono or dicycloalkylamino (3-8 carbon atoms in each ring), hydroxyrne h l , acyl of the formula -C(0)R, cyano, alkylthio (1-4 carbon atoms), alk lsulfinyl (1-4 carbon atoms), alkylsulfonyl (1-4 carbon atoms), cycloalkylthio (3-8 carbon atoms), cycloaliylsulfinyi (3-8 carbon atoms), cycloalkylsulfonyl (3-8 carbon atoms) , sulfamoyl , mono or dialkylsulfamoyl (1-4 carbon atoms in each alkyl group) , mono or dicycloalkylsulfamoyl (3-8 carbon atoms in each ring), mercapto, carboxy, carbamoyl (-C(0)-NH2), mono or dialkylcarbamoyl (1-4 carbon atoms in each alkyl group), mono or dicycloalkyl- carbamoyl (3-8 carbon atoms in each ring), alkoxycarbonyl (1-4 carbon atoms in the alkyl moiety), cycloalkoxy- carbonyl (3-8 carbon atoms in the ring), alkenyl (2-4 carbon atoms), cycloalkenyl (4-8 carbon atoms), alkynyl (2-4 carbon atoms), or two R2 taken together on contiguous carbon atoms can form a carbocyclic ring of 5-7 members; -347- 112249/7 = 0-33 wherein Ax is phenyl, tliienyl, furanyl, pynolyl, pyriinidyl, imidazolyl, ' pyrazmyl, oxazoly), i¾azojy3, naphibyl, benzofayl beiizofuranyl, indolyl, quinolmyl, isoquinolinyl and quimzolinyl; at least one of A-E cannot be carbon, unless R3 and R4 taken together form an aromatic or heteroaro- matic ring, as defined below; any one of the A-E can be N -with proviso that if R3 and R4 are either a lone pair on N or H, then n=0 Or 1; both B and E can be N, with the following provisos: the other two atoms of A-E in that ring are C, R4 is not- H, and n is 0 or 1; both A and D can be N, with the following provisos: the other two atoms of B-C in that ring are C, R3 is not H, and n is 0 or 1 ; any two contiguous positions in A-E can be a single heteroatom, N, O or S, wherein one of the two remaining named positions of A-E must be C{H), and the other can be C(H) or N, resulting in a 5 membered fused ring; R3 and R4 are independently H, alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy {1-4 carbon atoms) , cycloalkoxy (3-8 carbon atoms), hydroxy, acyloxy (1-4 carbon atoms), amino, ' mono or dialkylamino (1-4 carbon atoms) , mono or dicycloalkylamino (3-8 carbon atoms), carbonato (-OC(O)OR), where R is alkyl (1-4 carbon atoms) or cycloalkyl (3-8 carbon atoms), mono or dialkyl substituted ureido, thioureido, or N- or O-linked urethano (1-4 carbon atoms in each alkyl group), alkylthio (1-4 carbon atoms) or cycloalkylthio (3-8 carbon atoms), mercapto, alkenyl (2-4 carbon atoms), hydrazine ' -lower alkylhydrazino (1-4 carbon atoms) , acylamino (1-4 carbon atoms) , hydroxy1amino, . O-alkylhydroxylamino (1-4 carbon atoms) , or taken together can be methylene-, ethylene- or propylenedioxy, or taken together form a fused pyrrolidine, tetrahydrofuran, piperidine, piperazine, morpholine or thiomorpholine ring, or taken - 147a - together form a further aromatic ring selected from the group consisting of phenyl, furan, thiophene, pyrazole, isoxazole, isothiazole, pyrrole, oxazole, imidazole and thiazole, optionally substituted with 1-3 of the R2 substituents ; if B and D are C, A and E can be N, provided that at least one of R3 and R4 is lower alkoxy, or the two taken together form an aromatic ring selected from the group consisting of phenyl, furan, thiophene, pyrrole, oxazole, imidazole and thiazole, optionally substituted with 1-3 of the R2 substituents; and if one or two of A through E are N, then if R3 or R4 is on a neighboring C atom to one of the N atoms, the named substituent cannot be either OH or SH; 112249/9 R5 and R6 are H when A and/orE are a carbon atom; if any of the substitutents R1, E2, R3 0rR4 contain chiral centers, or in the case wherein R1 creates chiral centers on the linking atoms, then all stereoisomers thereof both separately and as racemic and/or diastereoisomeric mixtures are included- 2- A pharmaceutical composition according "to Claim 1, adapted for inhibiting Erb-B2 or Erb-B3 or Erb-B4 receptor tyrosine kinase comprising an effective inhibiting amount of a compound of Formula I . as defined in Claim 1; or a pharmaceutical salt or hydrate thereof -3 _ A pharmaceutical composition according to Claim .1 adapted for treating cancer comprising an effective cancer inhibiting amount of a compound of Formula I as defined in Claim 1; or a pharmaceutical sale or hydrate thereo -4- A pharmaceutical composition according to Claim 1 adapted for treating psoriasis comprising an effective psoriasis inhibiting amount of a compound of Formula J- as defined in Claim 1; or a pharmaceutical salt D hydrate thereof-5 - A pharmaceutical composition according to Claim 1, adapted for preventing blastocyte implantation comprising an effective blastocyte implantation inhibiting amount of a compound of Formula I . as defined in Claim 1; or a pharmaceutical salt or hydrate thereof. 6. A contraceptive composition according to Claim 1, comprising a contraceptively effective amount of a - 149 - compound of Formula I as defined in Claim 1 , or a pharmaceutical salt or hydrate thereof in admixture with a contraceptively acceptable excipient, diluent or carrier. 7. A pharmaceutical composition according to Claim 1, adapted for treating kidney disease comprising an effective kidney disease inhibiting amount of a compound of Formula I as defined in Claim 1; or a pharmaceutical salt or hydrate thereof - 8. A pharmaceutical composition according to Claim 1, adapted for treating pancreatitis comprising an effective amount of a compound of Formula I as defined in Claim 1; or a pharmaceutical salt or hydrate thereof - 9. A compound of Formula I : Formula I where : X = 0, S or NH n = 0, 1, 2; R1 = H or alkyl (1-4 carbon atoms); if n = 2 , R1 can be independently H or alkyl (1-4 carbon atoms) on either linking carbon atom; " l JV ' 112249/6 R2 is alkyl (1-4 carbon atoms), cycloalkyl (3-8 carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms), halo (fluoro, cbloro, bromO, iodo), perfluoroalkyl (1- carbon atoms), hydroxy, acyloxy of the formula; -0-C(O)R where E is alkyl of 1-carbon atoms or cycloalkyl of 3-8 carbon atoms, amino, mono or dialkylaminb (1-4 carbon atoms in each alkyl group), mono or dicycloalkyla ino (3-8 carbon atoms in each ring), hydroxy ethyl , acyl of the formula -C(0)R, cyano, alkylthio (1-4 carbon atoms), alkylsulfinyl (1-4 carbon atoms), alkylsulfonyl (1-4 carbon atoms), cycloaikylthio (3-8 carbon atoms), cycloalkyisulfml (3-8 carbon atoms), cycloalkylsulionyl (3-8 carbon atoms), sulfamoyl, mono or dialkylsuliaraoyl (1-4 carbon atoms in each alkyl group), mono ox dicycloalkylsul-.amoyl (3-8 carbon atoms in each ring), mercapto carboxy, carbamoyl (-C(O)-NH-,), mono or diaIky1carbamoyl (1-4 carbon atoms in each alkyl group), mono or dicycloalkyl-carbamoyl (3-8 carbon atoms in each ring) , alkoxycarbonyl (1-4 carbon atoms in the alkyl moiety), cycloaikoxy-carbonyl (3-8 carbon atoms in the ring) , ' alkenyl (2-4 carbon atoms), cycloalkenyl (4-8 carbon atoms), alkynyl (2-4 carbon atoms), or two R2 taken together on contiguous carbon atoms can form a carbocyclic ring of 5-7 members; and m = 0-3, wherein Ar is phenyl, thienyl, furanyl, pyrrolyl , pyrimidyl, imidazolyl, pyrazinyl, oxazolyl, thiazolyl, naphthyl, benzothienyl , benzo- furanyl, indolyl, quinolinyl, isoquinolinyl and quina- zolinyl ; at least one of A-E cannot be carbon, unless R3 and R" taken together form an aromatic or heteroaro-matic ring, as defined below; any one of the A-E can be N with proviso that if R3 and R* are either a lone pair on N or H, then n=0 or 1 ; - 150a - 112249/3 both B and E can be N, with the following PrOV4iSCS: he other two atoms of A-E in that ring are C, R4 is not H, and n is 0 or i; both A and D can be N, with the following provisos the other two atoms of B-C in that ring are C, R3 is not H, and n is 0 or i; R3 and R4 are independently H, alkyl (1-4 carbon atoms), cycloalkyl (3-a carbon atoms), alkoxy (1-4 carbon atoms), cycloalkoxy (3-8 carbon atoms) , hydroxy, acyloxy (1-4 carbon atoms) , amino, mono or dialkylamino (1-4 carbon atoms) , mono or dicycloalkylamino (3-8 carbon atoms) , carbonato (-OC(O)OR), where R is alkyl (1-4 carbon atoms) or cycloalkyl (3-8 carbon atoms), mono or dialkyi substituted ureido, thioureido, or N- or O-linked urethano (1-4 carbon atoms in each alkyl group , alkylthio (1-4 carbon atoms) or cycloalkylthio (3-8 carbon atoms), mercapto, alkenyl (2-4 carbon atoms), hydrazine N' -alkylhydrazirio '(1-4 carbon atoms), acylamino (1-4 carbon atoms) , hydroxy1amino, O-alkylhydroxylamino (1-4 carbon atoms), or taken together can be methylene-ethylene- or propyienedioxy, or taken together form a fused pyrrolidine, tetrahydrofuran, piperidine, piperazine, morpholine or thiomorpholine ring, or taken together form a further aromatic ring selected from the group consisting of phenyl, furan, thiophene, pyrazole, isoxazoie, isothiazole, pyrrole, oxazole, imidazole and thiazole, optionally substituted with 1-3 of the R2 substituents ; if B and D are C, A and E can be N, provided that at least one of R3 and R4 is lower alkoxy, or the two taken together form an aromatic ring selecced from the group consisting of phenyl, furan, thiophene, pyrrole, oxazole, imidazole and thiazole, optionally substituted with 1-3 of the R2 substituents; and -151- 112249/10 if one or two of A through E are N, then if R3 or is on a neighboring C atom to one cf the N atoms, the named substituent cannot be either OH or SB; and R5 and R6 are H when A and/or E are a carbon atom; if any of the substituted B2, R3 orR4 contain chiral centers, or in the esse wherein R creates chiral ^ centers on the linking etoras, then all stereoisomers thereof both separately and as racemic and/or diastereoisomeric mixtures are included/ - 152 - 112249/5 or a pharmaceutical salt or hydrate · thereof . 10.· The compound of claim 9, wherein the compound is selected from the group consisting of 6-Amino-4- ( 3-bromoanilino)pyrido[3 , 2-d]pyrimidine; 4- (3-Bromoanilino-6-methylaminopyrido[3 , 2~d]pyrimidine; 4- ( 3-Bromoanilino) - 6-dimethylaminopyrido[3 , 2-d]pyrimidine ; 7-Amino-4- ( 3-nitroan l no)pyrido[4 , 3-d ]pyrimidine ; 7-Amino-4- ( 3-bro oanilino) pyrido [4 , 3-d ]py imidine; 7-Amino-4- ( 4-bromoanilino) pyrido[ 4 , 3-d ]pyrimidine ; 7-Amino-4- ( 3-trif luoromethylanilino) pyrido[4 , 3-d]pyrididine ; 7-Acetamido-4-benzylaminopyrido- [ , 3-d] pyrimidine; 4- ( 3-Bromoanilino) -6-methylaminopyrido[ 3 ,4-d]pyrimidine; and 4- (3-Bromoanilino) -6-dimethylaminopyrido [ 3 , -d] pyrimidine . 11. A pharmaceutical composition adapted for inhibiting epidermal growth factor receptor tyrosine kinase comprising an effective inhibiting- amount of a compound of Formula I as defined in Claim 9, or a pharmaceutical salt or hydrate thereof. 12. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B , D & E carbon, with A nitrogen and R3 or R4 H, with the other one alkoxy or halogen. 13. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 o .h H, with the other one amino- 14. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case RJ - H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A - 153 - 112249/ 4 nitrogen and R3 or R4 H, with the other one mono or dialkylamino . 15. The composition of claim 11 wherein X = NH , n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 or R4 H, with the other one hydrazino. 16. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 or RA H, with the other one alkyl . 17. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 and R4 alkoxy. 18. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen and R3 and R4 alkyl. 19. The composition of claim 11 wherein X = NH , n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen, and R3 or RA amino, with the other one alkoxy. 20. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A nitrogen, and R3 or mono or dialkylamino, with the other one alkoxy. 21. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B , D & E carbon, with A nitrogen and R3 mono or dialkylamino, with RA hydroxy . 22. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, B, D & E carbon, with A - 154 - 112249/3 nitrogen, and R3 and R*1 taken together are roethylenedioxy, ethylenedioxy, 2 , 3-fused piperazine , 2,3-fused morpholine or 2,3-fused thiomorpholine . 23. The composition of claim 21 having any one of the following ring structures: 24. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and RA alkoxy or halogen - 25. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R^ amino. 26. The composition of claim 11 wherein X = NH , n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R4 mono or dialkylamino . 27.. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen and R^ hydrazino. 28. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring - 155 - 112249/ 4 phenyl optionally substituted, A, C & E carbon, with B nitrogen and R4 alkyl. 29. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 alkoxy or halogen. 30. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 amino. 31. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 mono or dialkylamino. 32. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 hydrazino. 33. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen and R3 alkyl . 34. The composition of claim 11 wherein X = NH , n = 0 or 1 , in which case R1 = H , the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or RA H, with the other one alkoxy. 35. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H , the aromatic ring phenyl optionally substituted, A, B &. D carbon, with E nitrogen and R3 or R4 H, with the other one amino. 36. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or Ri H, with the other one mono or dialkylamino. - 156 - 1 12249/4 37. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or R* H, with the other one hydrazino. 38. The composition of claim 11 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 or R4 H, with the other one alkyl. 39. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 and R4 alkoxy. 40. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R3 and Rh alkyl. 41. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen, and R3 or R4 amino, with the other one alkoxy. 42. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen, and R3 or R4 lower mono or dialkylamino , with the other one .· alkoxy. 43. The composition of -claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen and R4 mono or dialkylamino, with R3 hydroxy . 44. The composition of claim 11 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, A, B & D carbon, with E nitrogen, and R3 and R4 taken together are methylenedioxy, - 157 - 112249/4 ethylenedioxy, 2,3-fused piperazine, 2 , 3-fused morpholine or 2,3-fused thiomorpholine. 45. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 alkoxy. 46. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 mono or dialkylamino . 47. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 amino. 48. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, A & D carbon, with B and E nitrogen and R4 hydrazino. 49. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbon, with A and E nitrogen and R3 and R4 alkox . 50. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbon, with A and E nitrogen and R3 and R4 mono or dialkylamino. 51. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbon, with A and E nitrogen and R3 or R4 alkoxy, with the other mono or dialkylamino. 52. The composition of claim 11 wherein X = NH, n = 0, the aromatic ring phenyl optionally substituted, B & D carbon, with A and E nitrogen and R3 and R4 taken together are ethylenedioxy, 2,3-fused piperazine, 2,3- fused morpholine or 2,3-fused thiomorpholine. 53. The composition of claim 1 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a sulfur atom, with D & E carbon, or A & B - 158 - 112249/4 are carbon with D and E taken together as a sulfur atom, with R¾ or R3 H, alkyl, alkoxy, amino, or mono or dialkylamino. 54. The composition of claim 1 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are an oxygen atom, with D & E carbon, or A & B are carbon with D and E taken together as an oxygen atom, with R4 or R3 H, alkyl, alkoxy, amino, or mono or dialkylamino. 55. The composition of claim 1 wherein X = NH, n = 0 or 1 , in which case R3 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a nitrogen atom, with D & E carbon, or A & B are carbon with D and E taken together as a nitrogen atom, with R4 or R3 H, alkyl, alkoxy, amino, or : mono or dialkylamino. 56. The composition of claim 1 wherein X = NH , n = 0 or 1 , in which case R1 = H , the aromatic ring phenyl optionally substituted, and either A and B taken together are a sulfur atom with D carbon and E nitrogen, or D and E taken together are a sulfur atom, and A is nitrogen and B is carbon, with R3¾ H, alkyl, ■ alkoxy, amino, or mono or dialkylamino. 57. The composition of claim 1 wherein X = NH , n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are an oxygen atom with D carbon and E nitrogen, or D and E taken together are an oxygen atom, and A is nitrogen and B is carbon, with R3/i H, . alkyl, alkoxy, amino, or lower mono or dialkylamino. 58. The composition of claim 1 wherein X = NH, n = 0 or 1 , in which case R1 = H, the aromatic ring phenyl optionally substituted, A and B taken together are a nitrogen atom, and D is carbon and E is nitrogen, with - 159 - 11224 / 5 R3/6 H, or alkyl, and H, alkyl, alkoxy, amino, or . mono or dialkylamino. 59. The composition of claim 1 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are an oxygen atom with Ώ being nitrogen and E carbon, or B and D taken together are a nitrogen atom with A being nitrogen and E carbon, with R5/6 H, alkyl, alkoxy, amino, or mono or dialkyl-am no. 60. The composition of claim 1 wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a sulfur atom with D being nitrogen and E carbo , with R5/6 H, alkyl, alkoxy, amino, or mono or dialkyl- amino . 61. The composition of claim 1.. wherein X = NH, n = 0 or 1, in which case R1 = H, the aromatic ring phenyl optionally substituted, and either A and B taken together are a nitrogen atom with D being nitrogen and E carbon, or B and D taken together are a nitrogen atom with A being carbon and E nitrogen atom, with ^"/6 H or lower alkyl if on nitrogen, or H, alkyl, alkoxy, amino, or mono or dialkylamino if on carbon . 62. The composition of claim 11 wherein R1, R2, R3 or R¾ contain chiral centers, or in the case wherein R1 creates chiral centers on the linking atoms, then all stereoisomers thereof both separately and as racemic and/or diastereoisomeric mixtures are included therein. 63. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen, 1 H and R4 alkoxy. - 160 - 112249/ 4 64. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted. A, D & E carbon, with B nitrogen, R1 H and R4 amino or acylamino. 65. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen, R1 H and R4 mono or dialkylamino. 66. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, D &. E carbon, with B nitrogen, R1 H and R4 hydxazino. 67. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, D & E carbon, with B nitrogen, R1 H and R4 alkyl. 68. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen, R1 H and R3 alkoxy. 69. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted. A, B & E carbon, with D nitrogen, R1 H and 3 amino. 70. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen, R1 H and R3 mono or dialkylamino. 71. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, B & E carbon , with D nitrogen, R1 H and R3 hydrazino. 72. The composition of claim 11 wherein X=NH, n=0 or 1, the aromatic ring phenyl optionally substituted, A, B & E carbon, with D nitrogen, R1 H and 3 alkyl. 73. The composition of claim 11 having the ring structure below: - 161 - 112249/2 74. The composition of claim 11 having the ring structure below: 75. The composition of claim 11 having the ring structure below: 76. The composition of claim 11 having the ring structure below: 77. The composition of claim 11 having the ring structure below: - 162 - 112249/3 78 . The composition of claim 11 having the ring structure below: 79 . The composition of claim 1 having either of the ring structures below: Where 2 - nitrogen, oxygen or sulfur 80 . The composition of claim 1 wherein the compound is selected from the group consisting of 6-Amino-4- (3-bromoanilino)pyrido[3,2-d]pyrimidine; 4 - ( 3 - Bromoanilino) -6-methylaminopyrido[3 , 2-d]pyrimidine 4- ( ?- Bromoani1ino) -6-dimethylaminopyrido[ 3 , 2-d ] yrimidine ; 7- Amino-4- ( 3-nitroanilino)pyrido[4 , 3-d]pyrimidine ; 7-Amino- 4- ( 3-bromoanilino) pyrido[ 4 , 3-d ]pyrimidine ; 7-Amino-4- ( 4- bromoanilino ) pyrido [4 , 3-d] yrimidine; 7-Amino-4- ( 3- 112249/3 trif luoromethylanilino ) pyr ido [ 4 , 3-d] pyrimidine ; 7 Acetylamino-4- ( 3 -bromoanil no ) pyr ido [ 4 , 3-d ] pyrimidine 4-Benzy laminopyr ido [ 4 , 3-d ] yrimidin ; 7-Acetamido-4 benzyl aminopyr ido [ 4 , 3-d] pyrimidine ; 4- ( 3 -Bromoanil no ) -6 chloropyrido[3 , 4-d] yrimidine; 4- ( 3 -Bromoanilino ) -6 methoxypyrido[3 , 4-d] pyrimidine ; 4- ( 3-Bromoanilino ) -6 methyl aminopyr ido [ 3 , 4-d ] pyrimidine ; 4- ( 3-Bromoanilino ) -δ diroethylaminopyrido[3,4-d]pyrimidine; 4-(3 Bromoanilino) thieno[3 , 2-d]pyrimidine ; 4 Benzylaminothieno[3 , 2-d ] yrimidine ; 4- (3 Bromoanilino)thieno[2,3-d]pyrimidine; and N6- ( 3-Bromo phenyl ) adenine . 96i.28-4-Clainis-.\'0 be-23.11. :997